Alomone Labs has developed two innovative platforms for studying protein-protein interaction and/or protein co-localization in complex immunostaining or immunoprecipitation protocols:
ATTO-Conjugated Antibodies These antibodies can be used in immunohistochemistry (IHC) and immunocytochemistry (ICC) with same species antibodies.
Save time and resources with ONE antibody.
Benefits of immunostaining with ATTO-conjugated antibodies: ✓ Direct immuno-fluorescence ✓ Secondary antibodies are not necessary ✓ Ideal for immuno-colocalization with same species/host antibodies
Guinea Pig-Raised Antibodies Antibodies raised in guinea pig can be used with any non-guinea pig second species antibodies (e.g. antibodies raised in mouse, rabbit, goat, etc.) in complex immunostaining or immunoprecipitation protocols. Our line of guinea pig raised antibodies can be combined with any of our rabbit-raised or ATTO-conjugated antibodies in complex immunostaining protocols.
Immuno-Colocalization of TRPV4 and Aquaporin 2 in Mouse Kidney
Immunohistochemical staining of mouse kidney sections using Anti-TRPV4 Antibody (#ACC-034) and Anti-Aquaporin 2-ATTO-590 Antibody (#AQP-002-AO). TRPV4 staining (green) is detected in the tubules of the cortex and meddula (left panel). AQP2 immunoreactivity (red) is detected in the same areas (middle panel). Merged image shows complete overlap between TRPV4 and AQP2 staining.Adapted from Berrout, J.et al. (2012) J. Biol. Chem.287, 8782. with permission of the American Society for Biochemistry and Molecular Biology.
Immuno-Colocalization of VGLUT2 and Neurokinin 1 Receptor in Rat DRG
Immunohistochemical staining of perfusion-fixed frozen rat dorsal root ganglion (DRG) sections using Anti-VGLUT2-ATTO-594 Antibody (#AGC-036-AR), (1:60) and Anti-Neurokinin 1 Receptor (NK1R) (extracellular)-ATTO-488 Antibody (#ATR-001-AG), (1:60). A. VGLUT2 staining (red). B. NK1 receptor staining (green). C. Merge of the two images demonstrates co-localization in some neuronal bodies (arrows point at examples). Cell nuclei are stained with DAPI (blue).
Immuno-Colocalization of KCa3.1 and P2Y2 in Human Ovarian Carcinoma
Immunohistochemical staining of human ovarian carcinoma biopsies using Mouse Anti-KCNN4 (KCa3.1, SK4) (extracellular) Antibody (#ALM-051) and rabbit Anti-P2Y2 Receptor Antibody (#APR-010). Positive co-expression is detected in ovarian neoplastic cells (merged image). Expression of KCa3.1 is also observed in stromal cells (*); however, P2Y2 does not co-localize with KCa3.1 in these cells.Adapted from Robles-Martinez, L.et al. (2017) Sci. Rep. 7, 4340. with permission of Nature Publishing Group.
Immuno-Colocalization of GluN1 and ASCT1 in Mouse Hippocampus
Immunohistochemical staining of immersion-fixed, free floating rat brain frozen sections using Guinea pig Anti-NMDAR1 (GluN1) (extracellular) Antibody (#AGP-046), (1:600) and rabbit Anti-ASCT1 (SLC1A4) Antibody (#ANT-081), (1:300). A. GluN1 (in red) appears in profiles of hippocampal dentate gyrus interneurons (arrows). B. ASCT1 (in green) appears in the extracelullar matrix of the subgranular layer (SGL). C. Merge of the two images demonstrates adjacent localization in several neurons (arrows). Cell nuclei are stained with DAPI (blue). GL = granule layer.
Immuno-Colocalization of GIRK2 and LRRK2 in Mouse Substantia Nigra
Immunohistochemical staining of immersion-fixed, free floating mouse brain frozen sections using Guinea pig Anti-GIRK2 (Kir3.2) Antibody (#AGP-013), (1:200) and rabbit Anti-LRRK2 Antibody(#ANR-102), (1:200). A. Kir3.2 staining (red) appears in profiles of dopaminergic neurons (arrows). B. LRRK2 staining (green) is detected in several cell bodies in the SNC. C. Merge of the two images demonstrates colocalization in several neurons (arrows). Cell nuclei are stained with DAPI (blue).
Immuno-Colocalization of GluN2A and SynGAP in Rat Cingulate Cortex
Immunohistochemical staining of immersion-fixed, free floating rat brain frozen sections using Anti-NMDAR2A (GluN2A) (extracellular)-ATTO-488 Antibody(#AGC-002-AG), (1:200) and Anti-SynGAP Antibody (#APZ-032), (1:200), followed by donkey-anti-rabbit-Cy3. A. GluN2A staining (green) appears in neuronal profiles (arrows). B. SynGAP staining (red) is detected mostly in apical dendrites (horizontal arrows) an in neuronal soma (vertical arrows). C. Merge of the two images demonstrates co-localization in some neurons (vertical arrows). Cell nuclei are stained with DAPI (blue).
Immuno-Colocalization of Kir6.2 and BDNF in Mouse Hippocampus
Immunohistochemical staining of immersion-fixed,free floating mouse brain frozen sections using Guinea pig Anti-Kir6.2 Antibody (#AGP-067), (1:300) and Anti-BDNF Antibody (#ANT-010), (1:300). A. Kir6.2 staining (red) appears in the dentate gyrus granule layer (G) and in hilar interneurons (arrows). B. BDNF staining (green) in the same section appears in the dentate gyrus granule layer (G) and in hilar interneurons (arrows). C. Merge of the two images reveals colocalization of Kir6.2 and BDNF in hippocampal interneurons.
Immuno-Colocalization of Aquaporin 4 and TrkB in Rat Cerebellum
Immunohistochemical staining of perfusion-fixed frozen rat cerebellum sections using Anti-TrkB (extracellular)-ATTO-488 Antibody (#ANT-019-AG), (1:80) and Anti-Aquaporin 4 (AQP4) (300-314)-ATTO-594 Antibody (#AQP-014-AR), (1:80). A. TrkB labeling (green) appears in Purkinje cells (arrows) forming the Purkinje layer (P). B. Aquaporin 4 staining (red) appears in outlines of the “baskets” formed around Purkinje cells (vertical arrows) and in some blood vessels (horizontal arrow). C. Merge of panels A and B demonstrates the localization of TrkB and Aquaporin 4 in adjacent cellular compartments of the cerebellum. Nuclei were demonstrated using DAPI as the counterstain (blue).
Immuno-Colocalization of Synapsin-2 and GABA(A) α1 Receptor in Rat Striatum
Immunohistochemical staining of immersion-fixed, free floating rat brain frozen sections using rabbit Anti-Synapsin II (SYN2) Antibody(#ANR-015), (1:300) and Guinea pig Anti-GABA(A) α1 Receptor (extracellular) Antibody (#AGP-083), (1:600). A. Synapsin-2 staining (red) appears in Islands of Calleja (IC), (arrow). B. GABA(A) α1 Receptor (green) is also detected in Islands of Calleja and in adjacent neurons. C. Merge of the two images demonstrates colocalization in IC while adjacent GABA(A) α1 positive neurons do not express Synapsin-2. Cell nuclei are stained with DAPI (blue).
Immuno-Colocalization of Slitrk1 and KV2.1 in Rat Parietal Cortex
Immunohistochemical staining of immersion-fixed, free floating rat brain frozen sections using rabbit Anti-Slitrk1 (extracellular) Antibody (#ASR-081), (1:200) and Guinea pig Anti-KV2.1 Antibody(#AGP-109), (1:200). A. Sltrk1 staining (red) appears in profiles of pyramidal neurons. B. KV2.1 staining (green) is detected in profiles of pyramidal neurons. C. Merge of the two images shows colocalization in several neurons (arrows). Cell nuclei are stained with DAPI (blue).
Immuno-Colocalization of VAChT and p75NTR in Rat Medial Septum
Immunohistochemical staining of immersion-fixed, free floating rat brain frozen sections using rabbit Anti-Vesicular Acetylcholine Transporter (VAChT) Antibody (#ACT-003), (1:200) and Mouse Anti-Rat p75 NGF Receptor (extracellular) Antibody (#AN-170), (1:200). A. VAChT staining (green) appears in several neuronal cells. B. p75NTR (red) also stains neuronal cells. C. Merge of the two images shows colocalization of VAChT and p75NTR in some cells (horizontal arrows), while other cells express only VAChT (vertical arrows). Cell nuclei are stained with DAPI (blue).
Immuno-Colocalization of Plexin-A2 and p75NTR in Rat Brain
Immunohistochemical staining of perfusion-fixed frozen rat brain sections using Anti-Plexin-A2 (extracellular) Antibody (#APR-082), (1:400) and Mouse Anti-Rat p75 NGF Receptor (extracellular) Antibody (#AN-170), (1:300). A. Sections of rat horizontal diagonal band (HDB) were stained for Plexin-A2 (green). B. Same sections were stained for p75NTR (red). C. Merge of the two images reveals cells expressing both Plexin-A2 and p75NTR (horizontal arrows), cells expressing only Plexin-A2 (upwards pointing arrows) or cells expressing only p75NTR (downwards pointing arrows) can also be observed. Cell nuclei are stained with DAPI (blue).
Immuno-Colocalization of Kir3.2 and KCNK2 in Rat Substantia Nigra
Immunohistochemical staining of immersion-fixed, free floating rat brain frozen sections using rabbit Anti-GIRK2 (Kir3.2) Antibody (#APC-006), (1:400) and Guinea pig Anti-KCNK2 (TREK-1) Antibody (#AGP-049), (1:120). A. Kir3.2 staining (red) appears in cells of the substantia nigra pars compacta (SNC, arrows). B. KCNK2 (green) appears in both compacta (SNC) and reticulata (SNR) portions of the substantia nigra. C. Merge of the two images reveals co-localization in some cells (arrows), mainly in the SNC region. Cell nuclei are stained with DAPI (blue).
Immuno-Colocalization of VGAT and Gephyrin in Rat Spinal Cord
Immunohistochemical staining of perfusion-fixed frozen rat spinal cord sections using Anti-Gephyrin Antibody (#AIP-005), (1:300) and Anti-Vesicular GABA Transporter (VGAT)-ATTO-594 Antibody (#AGT-005-AR), (1:80). A. Gephyrin staining (green) appears in the superficial layer of the spinal dorsal horn and some cellular profiles were stained. B. VGAT staining (red) appears in superficial layer and in cell profiles. C. Merge images of A and B reveals co-localization (arrows) in superficial layers and separate expression in some cells. Cell nuclei were stained with DAPI (blue) as the counterstain.
Immuno-Colocalization of Kir4.1 and α1-Syntrophin in Rat Fornix
Immunohistochemical staining of immersion-fixed, free floating rat brain frozen sections using Guinea pig Anti-Kir4.1 (KCNJ10) Antibody (#AGP-012), (1:400) and Anti-α1-Syntrophin (SNTA1) Antibody (#APZ-021), (1:300). A. Kir4.1 staining (red) appears in blood vessel profiles (arrows) in the fornix. B. Syntrophin alpha 1 staining in the same section (green) appears in several elements including blood vessels (arrows). C. Merge of the two images reveals partial co-localization of Kir4.1 and syntrophin alpha 1 in hilar cells. Cell nuclei are stained with DAPI (blue).
Immuno-Colocalization of proNGF and Cannabinoid Receptor 1 in Mouse Hippocampus
Immunohistochemical staining of immersion-fixed, free floating mouse brain frozen sections using Guinea pig Anti-proNGF Antibody (#AGP-031), (1:300) and rabbit Anti-Cannabinoid Receptor 1 (extracellular) Antibody (#ACR-001), (1:300). A. proNGF staining (red) appears in the dentate gyrus (DG) granule layer (G) and in hilar interneurons (arrows). B. CB1R staining (green) appears in axonal processes surrounding the granule layer and a few hilar interneurons (arrows). C. Merge of the two images reveals co-localization of proNGF and CB1R in a few hilar cells. Cell nuclei were stained with DAPI (blue).
Immuno-Colocalization of NET and NK1 Receptor in Rat Brain Stem
Immuno-Colocalization of GluR1 and PICK1 in Rat Hippocampus
Immunohistochemical staining of immersion-fixed, free floating rat brain frozen sections using rabbit Anti-PICK1 Antibody (#APZ-014), (1:400) and Guinea pig Anti-GluR1 (GluA1) (extracellular) Antibody (#AGP-009), (1:400). A. GluR1 staining (green). B. PICK1 staining (red) in the same section. C. Merge of the two images reveals co-localization in several interneurons (arrows) in the dentate gyrus (DG).
Immuno-Colocalization of TRPM8 and TrkA in Rat DRG
Immunohistochemical staining of perfusion-fixed frozen rat dorsal root ganglia (DRG) sections using Anti-TrkA (extracellular)-ATTO-633 Antibody (#ANT-018-FR), (1:60) and Anti-TRPM8 (extracellular) Antibody (#ACC-049), (1:300). A. TRPM8 labeling followed by goat-anti-rabbit-Alexa-488 (green). B. The same section was then labeled for TrkA (purple). C. Merge of A and B demonstrates co-localization of TRPM8 and TrkA in rat DRG (arrows). Cell nuclei were stained with DAPI (blue).
Immuno-Colocalization of ASIC1 and ASIC3 in Rat DRG
Immunohistochemical staining of perfusion-fixed frozen rat dorsal root ganglia (DRG) sections using Guinea pig Anti-ASIC1 Antibody (#AGP-053), (1:400) and rabbit Anti-ASIC3 Antibody (#ASC-018), (1:400). A. ASIC3 staining. B. ASIC1 staining in same section (green). C. Merge of the two images shows some co-localization (arrow). Cell nuclei were stained with DAPI (blue).
Immuno-Colocalization of DPP6 and KV4.2 in Rat Hippocampus
Immunohistochemical staining of immersion-fixed, free floating rat brain frozen sections using rabbit Anti-DPP6 (extracellular) Antibody (#APC-146), (1:400) and Guinea pig Anti-KV4.2 Antibody (#AGP-038), (1:1000). A. DPP6 (green) is expressed in the CA3 rat hippocampal region. B. KV4.2 (red) is expressed in the same region. C. Merge of the two images demonstrates partial colocalization in neurons of the CA3 region (arrows). Cell nuclei are stained with DAPI (blue).
Immuno-Colocalization of GluN2A and GluN2B in Mouse Deep Cerebellar Nucleus
Immunohistochemical staining of perfusion-fixed frozen mouse brain sections using Anti-NMDAR2B (GluN2B) (extracellular)-ATTO-594 Antibody (#AGC-003-AR), (1:60) and Anti-NMDAR2A (GluN2A) (extracellular) Antibody (#AGC-002), (1:200). A. Sections were incubated with Anti-NMDAR2A (GluN2A) (extracellular) Antibody, followed by goat anti-rabbit-Alexa-488 (green). B. The same sections were incubated with Anti-NMDAR2B (GluN2B) (extracellular)-ATTO-594 Antibody (red). C. Merge of A and B demonstrates the ubiquitous colocalization of the GluN2A and GluN2B subunits in cells with neuronal profiles in this nucleus. Arrows point at an example of NR2A and NR2B co-expression.
Immuno-Colocalization of NaV1.8 and Synaptophysin in Rat DRG
Immunohistochemical staining of rat DRG frozen section using Anti-NaV1.8 (SCN10A)-ATTO-594 Antibody (#ASC-016-AR) and Anti-Synaptophysin Antibody (#ANR-013). A. NaV1.8 staining (red). B. Synaptophysin staining (green). C. Merged image demonstrates a partial overlap in the distribution of NaV1.8 and Synaptophysin within the DRGs. DAPI is used as the counterstain (blue).
Immuno-Colocalization of HCN4 and HCN2 in Mouse Thalamus
Immunohistochemical staining of mouse thalamus frozen section using Guinea pig Anti-HCN4 Antibody (#AGP-004) and rabbit Anti-HCN2 Antibody (#APC-030). A. Staining of HCN4 (green) appears in the ventral posterior thalamic nucleus (VPL). B. In the same section as in A, staining of HCN2 (red) appears in the ventral posterior thalamic nucleus (VPL) and also in the reticular thalamic nucleus (RT). The area between these thalamic nuclei (star) is white matter and neither protein is expressed in that region. C. Merged images of A and B.
Immuno-Colocalization of KV1.3 and KV1.5 in Mouse Cerebellum
Immunohistochemical staining of mouse perfusion-fixed frozen brain sections using Anti-KV1.3 (KCNA3) (extracellular)-Biotin Antibody (#APC-101-B) (1:400), and Anti-KV1.5 (KCNA5)-ATTO-550 Antibody (#APC-004-AO), (1:60). A. KV1.3 staining (green) is detected in Bergmann glia soma and processes in the molecular layer (Mol) (arrow). B. Same section shows staining for KV1.5 (red). C. Merge of the two images suggests considerable co-localization in the soma of Bergmann glia. In the molecular layer, the distribution of KV1.5 is diffuse, unlike the discrete staining for KV1.3 in glial processes. Cell nuclei were visualized with DAPI (blue).
Immuno-Colocalization of TrkB and BDNF in Mouse Hippocampus
A. Fixed brain sections were incubated with human BDNF-Biotin (#B-250-B) followed by Streptavidin-Cy3. B. Same sections were incubated with Anti-TrkB (extracellular)-ATTO-488 Antibody (#ANT-019-AG). C. Merge of A and B reveals co-localization in the pyramidal layer (P). Outside the pyramidal layer, there are also cells with only BDNF expression (red, horizontal arrows) or only TrkB expression (green, vertical arrows).
Immuno-Colocalization of GluN1 and Plexin-A1 in Rat Olfactory Bulb
Immunohistochemical staining of immersion-fixed, free floating rat brain frozen sections using Guinea pig Anti-NMDAR1 (GluN1) (extracellular) Antibody (#AGP-046), (1:600) and rabbit Anti-Plexin-A1 (extracellular) Antibody (#APR-081), (1:400). A. GluN1 (green) is expressed in the glomeruli (horizontal arrows). B. Plexin A1 staining (red) in the same section, shows expression in the glomeruli (arrow) but also in other layers (vertical arrow) of the olfactory bulb. C. Merge of the two images show cases co-localization in the glomeruli vs. lack of co-localization in deeper layers of the olfactory bulb. Cell nuclei are stained with DAPI (blue).
Immuno-Colocalization of GluN1 and M4 Muscarinic Receptor in Mouse Parietal Cortex
Immunohistochemical staining of immersion-fixed, free floating mouse brain frozen sections using rabbit Anti-M4 Muscarinic Receptor Antibody (#AMR-004), (1:400) and Guinea pig Anti-NMDAR1 (GluN1) (extracellular) Antibody (#AGP-046), (1:1200). A. M4 Muscarinic receptor staining (green). B. GluN1 staining (red) in same section. C. Merge of the two images reveals several cells expressing both M4 Muscarinic Receptor and GluN1 (arrows). Cell nuclei were stained with DAPI (blue).
Immuno-Colocalization of CaV1.2 and GABA(A) α1 Receptor in Rat Hippocampus
Immunohistochemical staining of mouse and rat hippocampal dentate gyrus using Guinea pig Anti-CaV1.2 (CACNA1C) Antibody (#AGP-001) and Anti-GABA(A) α1 Receptor (extracellular)-ATTO-488 Antibody (#AGA-001-AG) in the same section. Both CaV1.2 (red) and GABA(A) α1 Receptor (green) are detected in neuron-shapep cells (arrows). Staining suggests partial colocalization between CaV1.2 and GABA(A) α1 Receptor in a sub-population of rat dentate gyrus neurons.
Immuno-Colocalization of TRPC6 and TRPC3 in Rat Cerebellum
Immunohistochemical staining of rat cerebellum frozen section using Guinea pig Anti-TRPC6 Antibody (#AGP-002) and rabbit Anti-TRPC3-ATTO-594 Antibody (#ACC-016-AR). A. TRPC6 staining (green) appears in molecular layer and in Purkinje cells. B. In the same section as in A, staining of TRPC3 (red) appears as well in both molecular layer and Purkinje cells. C. Merge images of A and B indicates co-localization in Purkinje cells and molecular layer.
Immuno-Colocalization of GluN1 and CALHM1 in Mouse Hippocampal CA1 Region
Immunohistochemical staining of perfusion-fixed frozen mouse brain sections using Anti-NMDAR1 (GluN1) (extracellular) Antibody (#AGC-001), (1:200) and Anti-CALHM1-ATTO-594 Antibody(#ACC-101-AR), (1:60). A. Sections were stained with Anti-NMDAR1 (GluN1) (extracellular) Antibody, followed by goat-anti-rabbit-Cy2 (green). Staining reveals expression in neurons of the pyramidal layer (an arrow points at the layer). B. The same section was incubated with Anti-CALHM1-ATTO-594 Antibody (red). C. Merge of the two images demonstrates colocalization of GluN1 and CALHM1 in pyramidal neurons.
Immuno-Colocalization of ASIC1 and Parvalbumin in Rat Hippocampus
Immunohistochemical staining of immersion-fixed, free floating rat brain frozen sections using Guinea pig Anti-ASIC1 Antibody (#AGP-053), (1:300). A. ASIC1 (green) is detected in the hilus of the dentate gyrus (arrows). B. The same section was stained with anti-parvalbumin (red). C. Merge of the two images shows partial colocalization (orange, see arrows). Cell nuclei in the same section were stained with DAPI (Blue).
Immuno-Colocalization of Neurexin 1α and GABA(A) ε Receptor in Mouse Cerebellum
Immuno-Colocalization of mGluR1 and TRPC3 in Mouse Cerebellum
Immunohistochemical staining of perfusion-fixed frozen mouse cerebellum sections using Anti-TRPC3-ATTO-594 Antibody (ACC-016-AR), (1:60) and Anti-mGluR1 (extracellular)-ATTO-488 Antibody (#AGC-006-AG), (1:60). A. TRPC3 staining (red). B. mGluR1 staining (green). C. Merge of the two images suggests extensive co-localization in Purkinje cells (vertical arrows). Note expression of mGluR1 in Purkinje dendrites (horizontal arrow) but not of TRPC3. Cell nuclei are stained with DAPI (blue).
Immuno-Colocalization of PSD-95 and GluN2B in Rat Brain
Immunohistochemical staining of perfusion-fixed frozen rat parietal cortex sections using Anti-PSD-95 Antibody (#APZ-009), (1:400) and Anti-NMDAR2B (GluN2B) (extracellular)-ATTO-594 Antibody (#AGC-003-AR), (1:60). A. PSD-95 staining (green). B. The same sections were stained for GluN2B (red). C. Merge of the two images shows several cells expressing both proteins (arrows point to several examples). Nuclei are stained with DAPI (blue).
Immuno-Colocalization of nAChR α4 and nAChR β2 in Mouse Brain
Immuno-Colocalization of VMAT2 and TRPC3 in Rat Brain
Immunohistochemical staining of perfusion-fixed frozen rat substancia nigra sections using Anti-VMAT2-ATTO-488 Antibody (#AMT-006-AG), (1:60) and Anti-TRPC3-ATTO-594 Antibody (#ACC-016-AR), (1:60). A. VMAT2 staining (green). B. The same section stained for TRPC3 (red). C. Merged image demonstrates the ubiquitous colocalization of VMAT2 and TRPC3 in cells of the substantia nigra pars compacta. Arrows point at examples of co-expression.
Immuno-Colocalization of AT2 Receptor and VGLUT2 in Rat Supraoptic Hypothalamic Nucleus
Immunohistochemical staining of perfusion-fixed frozen rat brain sections using Anti-Angiotensin II Receptor Type-2 (extracellular)-ATTO-488 Antibody (#AAR-012-AG), (1:60) and Anti-VGLUT2-ATTO-594 Antibody (#AGC-036-AR), (1:60). A. AT2 Receptor staining (green). B. VGLUT2 staining (red). C. Merge of the two images shows a general lack of co-localization of AT2 and VGLUT2 in this part of the hypothalamus (arrow). Cell nuclei are stained with DAPI (blue).
Immuno-Colocalization of Aquaporin 2 and Neutrophil Gelatinase-Associated Lipocalin (Ngal) in Mouse Kidney
Immunohistochemical staining of mouse kidney sections using Anti-Aquaporin 2-ATTO-550 Antibody (#AQP-002-AO), (red) and anti-Ngal antibody (green). Some co-localization between the two proteins is observed in the medulla where AQP2 is expressed on the medulla side of cells.Adapted from Hains, D.S.et al. (2014) Am. J. Physiol.307, F869. with permission of the American Physiological Society.
Immuno-Colocalization of Aquaporin 2 and TRPV4 in Rat Kidney
Immunohistochemical staining of rat kidney sections using Anti-TRPV4 Antibody (#ACC-034) and Anti-Aquaporin 2-ATTO-550 Antibody(AQP-002-AO). Shown are Representative confocal plane micrographs (axes are shown) and corresponding cross-sections (pointed by arrows) showing three-dimensional stacks of TRPV4 (green), AQP2 localization (red), and the combined image localization (yellow). A. Wild-type Sprague-Dawley (S/D) rat. B. Cyst monolayer from a PCK453 rat. DAPI staining is shown in blue.Adapted from Zaika, O.et al. (2013) J. Am. Soc. Nephrol.24, 604. with permission of the American Society of Nephrology.
Immuno-Colocalization of PTH2R and VGLUT2 in Mouse Hippocampus
Immunohistochemical staining of mouse hippocampal CA3 region frozen sections using Anti-PTH2R (extracellular)-ATTO-488 Antibody (#APR-052-AG), (1:60) and Anti-VGLUT2-ATTO-594 Antibody (#AGC-036-AR), (1:60). A. PTH2 receptor staining (green). B. VGLUT2 staining (red) in same section. C. Merge of A and B suggests extensive localization of PTH2R on glutamatergic neurons.
Tityustoxin-Kα-ATTO-594 Colocalizes with KV1.2 Channel and Parvalbumin in Mouse Cerebellum
A. Staining of free-floating mouse brain sections using Tityustoxin-Kα-ATTO-594 (#STT-360-AR) (red). Sections were then stained using Anti-KV1.2 (KCNA2) Antibody(#APC-010), (1:200) followed by goat anti-rabbit-AlexaFluor-488 (green). Tityustoxin-Kα-ATTO-594 and KV1.2 are both detected in the pinceau structures (yellow staining, arrow) of the cerebellum. B. Same brain sections as in A were labeled with Tityustoxin-Kα-ATTO-594 (red) followed by anti-Parvalbumin. Tityustoxin-Kα-ATTO-594 (red) labeled the pinceau structures (arrow) of the cerebellum. Parvalbumin stained not only the pinceau but also the soma of Purkinje cells. Colococalization between the two is depicted in yellow.
Immuno-Colocalization of p75NTR and TrkA in Rat DRG
Immunohistochemical staining of perfusion-fixed frozen rat dorsal root ganglia (DRG) sections using Anti-p75 NGF Receptor (extracellular)-ATTO-488 Antibody (#ANT-007-AG), (1:60) and Anti-TrkA (extracellular)-ATTO-633 Antibody (#ANT-018-FR), (1:60). A. p75NTR staining (green). B. TrkA staining in same section (purple). C. Merge of the two images suggests extensive co-localization (arrow) between TrkA and p75NTR receptors in rat DRG. Cell nuclei were stained with DAPI (blue).
Immuno-Colocalization of CaV1.2 and GABA(A) α1 Receptor in Mouse Hippocampus
Immunohistochemical staining of mouse hippocampal dentate gyrus using Guinea pig Anti-CaV1.2 (CACNA1C) Antibody (#AGP-001) and Anti-GABA(A) α1 Receptor (extracellular)-ATTO-488 Antibody (#AGA-001-AG) in the same section. Both CaV1.2 (red) and GABA(A) α1 Receptor (green) are detected in neuron-shapep cells (arrows). Staining suggests partial colocalization between CaV1.2 and GABA(A) α1 Receptor in a sub-population of rat dentate gyrus neurons.
Immuno-Colocalization of VGLUT2 and P2X7 Receptor in Rat Spinal Cord
Immunohistochemical staining of perfusion-fixed frozen rat spinal cord sections using Anti-VGLUT2 Antibody (#AGC-036), (1:600) and Anti-P2X7 Receptor-ATTO-550 Antibody (#APR-004-AO), (1:100). A. Vesicular Glutamate Transporter 2 labeling followed by goat-anti-rabbit-Alexa-488 (green). B. The same section labeled for P2X7 Receptor (orange). C. Merge of A and B demonstrates partial co-localization of VGLUT2 and P2X7 Receptor in dorsal horn and in lateral column (L. Col., arrow). Cell nuclei were stained with DAPI (blue).
Immuno-Colocalization of KCa2.3 (SK3) and Aquaporin 2 in Mouse Kidney
Immunohistochemical staining of mouse kidney sections using Anti-KCNN3 (KCa2.3, SK3) (N-term) Antibody (#APC-025) and Anti-Aquaporin 2-ATTO-550 Antibody (#AQP-002-AO). A. Aquaporin 2 staining (red). B. SK3 staining (green). C. Merge of A and B shows SK3 expression in Aquaporin 2 positive tubules (orange).Adapted from Berrout, J.et al. (2014) PLoS ONE9, e95149. with kind permission of Pro. O'Neil, R.G., Dpt. or Integrative Biology, the University of Texas Health Science Center Medical School, Houston, Texas, U.S.A.
Immuno-Colocalization of KV1.1 and KV1.2 in Mouse Cerebellum
Immunohistochemical staining of mouse perfusion-fixed frozen brain sections using Anti-KV1.2 (KCNA2) Antibody (#APC-010), (1:300) and Anti-KV1.1 (KCNA1) (extracellular)-ATTO-594 Antibody (#APC-161-AR), (1:100). A. KV1.2 staining, followed by donkey-anti-rabbit-Cy2 (green). B. KV1.1 staining (red). C. Merge of the two images suggests considerable co-localization in the pinceau structures (up-pointing arrows). KV1.1 also appears in blood vessels (down-pointing arrows), where no KV1.2 expression is observed.
Immuno-Colocalization of CaV1.2 and GABA(A) α1 Receptor in Rat Cerebellum
Immunohistochemical staining of rat cerebellum using Guinea pig Anti-CaV1.2 (CACNA1C) Antibody (#AGP-001) and Anti-GABA(A) α1 Receptor (extracellular)-ATTO-488 Antibody (#AGA-001-AG). A. CaV1.2 (red) is detected mostly in Purkinje cells (arrow). B. In the same section, GABA(A) α1 Receptor (green) is observed in the granule layer. C. Merge of the two images suggests some colocalization between CaV1.2 and GABA(A) α1 Receptor in the rat granule layer but only CaV1.2 appears in Purkinje cells.
Immuno-Colocalization of TrkB in Mouse Cerebellum
A. Fixed brain sections were incubated with human BDNF-Biotin (#B-250-B) followed by Streptavidin-Cy3. B. Same sections were incubated with Anti-TrkB (extracellular)-ATTO-488 Antibody (#ANT-019-AG). C. Merge of A and B reveals co-localization in the Purkinje layer (P). TrkB positive cells (green) with BDNF binding (red, arrow) is seen.
Immuno-Colocalization of Melatonin Receptor Type 1B and AT2 Receptor in Rat Brain
Immunohistochemical staining of perfusion-fixed frozen brain sections using Anti-Melatonin Receptor 1B (MTNR1B) Antibody (#AMR-032), (1:600) and Anti-Angiotensin II Receptor Type-2 (extracellular)-ATTO-488 Antibody (#AAR-012-AG), (1:100). A. MTNR1B staining (red) (arrows). B. The same section labeled for Angiotensin II Receptor Type-2 (green). C. Merge of the two images suggests considerable co-localization in the paraventricular nucleus (arrows). For orientation, note localization with respect to 3rd ventricle (3rd V).
Immuno-Colocalization of Melatonin Receptor Type 1B and SCN2A (NaV1.2) in Rat Paraventricular Nucleus
Immunohistochemical staining of perfusion-fixed frozen rat paraventricular nucleus sections using Anti-Melatonin Receptor 1B (MTNR1B) Antibody (#AMR-032), (1:600) and Guinea pig Anti-SCN2A (NaV1.2) Antibody (#AGP-026), (1:2000). A. MTNR1B staining (red) (arrows). B. The same section labeled for NaV1.2 (green). C. Merge of A and B demonstrates partial co-localization of MTNR1B and NaV1.2 in the paraventricular nucleus (PVN). For orientation, note location with respect to the third ventricle (3rd V). Cell nuclei were stained with DAPI (blue).
Immuno-Colocalization of TrKA and NGF in Mouse Cerebellar Cortex
A. Fixed brain sections were incubated with mouse NGF 2.5S-Biotin (#N-240-B) followed by streptavidin DyLight-488 (green). Staining of Bergmann glial processes (vertical arrow) in the molecular layer (Mol) and of Purkinje cell soma (horizontal arrow). B.Same sections were incubated with Anti-TrKA (extracellular)-ATTO-633 Antibody (#ANT-018-FR). C. Merged image of A and B shows co-localization of NGF and TrKA mostly in the Purkinje layer (horizontal arrow). Nissl staining is used as the counterstain (blue).
Immuno-Colocalization of KCa3.1 and TRPC1 in Mouse Brain
Immunohistochemical staining of mouse brain sections using Mouse Anti-KCNN4 (KCa3.1, SK4) (extracellular) Antibody (#ALM-051) and Anti-TRPC1 Antibody (#ACC-010). YFP denotes neuroblasts. TRPC1 staining (blue) is detected in neuroblasts and outside neuroblasts as well (in astrocytes). KCa3.1 (red) is detected in neuroblasts. Merged image of all three staining (lower right panel) demonstrates the partial co-localization between the three.Adapted from Turner, K.L. and Sontheimer, H. (2014) Cereb. Cortex24, 2388. with permission of Oxford University Press.
Immuno-Colocalization of Kir4.1 and GFAP in Rat Hippocampal Dentate Gyrus
Immunohistochemical staining of rat frozen brain sections using Guinea pig Anti-Kir4.1 (KCNJ10) Antibody (#AGP-012), (1:300). A. Kir4.1 staining (green) reveals Kir4.1 clusters in the molecular layer (Mol). B. The same section was stained with mouse anti-glial fibrillary acidic protein (GFAP), (red) showing astrocyte profiles. C. Merge of Kir4.1 and GFAP images reveals colocalization on some branches of astrocytic fibers. In addition, Kir4.1 is expressed in the fine ramification of astrocytic fibers, thus creating a “halo-like” cloud around the GFAP astrocytic outline. DAPI (blue) counterstain reveals the outline of the dentate granule layer.
Immuno-Colocalization of VGLUT2 and Neurokinin 1 Receptor in Rat DRG
Immunohistochemical staining of perfusion-fixed frozen rat dorsal root ganglion (DRG) sections using Anti-VGLUT2-ATTO-594 Antibody (#AGC-036-AR), (1:60) and Anti-Neurokinin 1 Receptor (NK1R) (extracellular)-ATTO-488 Antibody (#ATR-001-AG), (1:60). A. VGLUT2 staining (red). B. NK1 Receptor staining (green). C. Merge of the two images demonstrates co-localization in some neuronal bodies (arrows point at examples). Cell nuclei are stained with DAPI (blue).
Immuno-Colocalization of TRPV1 and NaV1.8 in Rat DRG
Immunohistochemical staining of rat dorsal root ganglion (DRG) using Anti-Rat TRPV1 (VR1) (extracellular)-ATTO-488 Antibody (#ACC-029-AG), (green), (1:60) and Anti-NaV1.8 (SCN10A)-ATTO-594 Antibody (#ASC-016-AR), (red), (1:60). A. TRPV1 staining. B. NaV1.8 staining. C. Merge of A and B demonstrates partial co-localization of TRPV1 and NaV1.8 channels. Nuclei stained using DAPI as the counterstain (blue).
Immuno-Colocalization of TRPV1 and mGluR5 in Rat DRG
Immunohistochemical staining of rat dorsal root ganglion using Anti-Rat TRPV1 (VR1) (extracellular)-ATTO-488 Antibody (#ACC-029-AG), (1:60) and Anti-mGluR5 (extracellular)-ATTO-594 Antibody (#AGC-007-AR), (1:60). A. TRPV1 staining (green). B. mGluR5 staining of the same section (red). C. Merge of A and B demonstrates co-localization of TRPV1 and mGluR5 in DRG cells. Nuclei were stained using DAPI as the counterstain (blue).
Immuno-Colocalization of NK1 Receptor and mGluR5 in Rat DRG
Immunohistochemical staining of rat dorsal root ganglion using Anti-Neurokinin 1 Receptor (NK1R) (extracellular)-ATTO-488 Antibody (#ATR-001-AG), (green), (1:60) and Anti-mGluR5 (extracellular)-ATTO-594 Antibody (#AGC-007-AR), (red), (1:60). A. NK1 receptor staining. B. mGluR5 staining appears in both cells (horizontal arrow) and fibers (vertical arrow). C. Merge of A and B demonstrates the co-localization of NK1 and mGluR5 receptors. Nuclei staining using DAPI as the counterstain (blue).
Immuno-Colocalization of TRPV2 and mGluR5 in Rat DRG
Immunohistochemistry of rat dorsal root ganglion using Guinea pig Anti-TRPV2 (VRL1) (extracellular) Antibody (#AGP-033) (1:60) and Anti-mGluR5 (extracellular)-ATTO-594 Antibody (#AGC-007-AR), (red), (1:60). A. TRPV2 staining (green). B. mGluR5 staining (red). C. Merge of A and B demonstrates co-localization of TRPV2 and mGluR5 in DRG cells. Nuclei are stained using DAPI as the counterstain (blue).
Immuno-Colocalization of Orexin Receptor 1 and Parathyroid Hormone 1 Receptor in Rat Ventromedial Hypothalamus
Immunohistochemical staining of perfusion-fixed frozen rat brain sections using Anti-Orexin Receptor 1-ATTO-488 Antibody (#AOR-001-AG), (1:60) and Anti-PTH1R (extracellular) Antibody (#APR-051), (1:60). A. Sections were incubated with Anti-PTH1R (extracellular) Antibody, followed by donkey-anti-rabbit-Cy3 (red). B. The same sections were incubated with Anti-Orexin Receptor 1-ATTO-488 Antibody, (green). C. Merge of A and B shows staining of Orexin receptor 1 in nerve fibers (vertical arrows) while Parathyroid hormone 1 receptor is apparent on cell bodies (horizontal arrows). Cell nuclei are stained with DAPI (blue).
Immuno-Colocalization of GluR1 and Vesicular GABA Transporter in Human U-87 MG Cells
Immunocytochemical staining of human glioblastoma U-87 MG. Extracellular staining of live intact cells with Guinea pig Anti-GluR1 (GluA1) (extracellular) Antibody (#AGP-009), (1:25), followed by goat anti-guinea pig-AlexaFluor-488 secondary antibody (green). Cells were subsequently fixed, permeabilized and labeled with Anti-Vesicular GABA Transporter (VGAT) Antibody (#AGT-005), (1:200), followed by goat anti-rabbit-AlexaFluor-594 secondary antibody (red). Representative merged images of the double labeled cells are shown in A and B.
Immuno-Colocalization of TRPC6 and TRPC3 in Rat Cerebellum
Immunohistochemical staining of rat cerebellum frozen section using Guinea pig Anti-TRPC6 Antibody (#AGP-002) and rabbit Anti-TRPC3 Antibody (#ACC-016). A. TRPC6 staining (green) appears in molecular layer and in Purkinje cells. B. In the same section, staining of TRPC3 (red) appears as well in both molecular layer and Purkinje cells. C. Merge images of A and B indicates extensive co-localization. DAPI is used as the counterstain (blue).
Immuno-Colocalization of P2X7 Receptor and Iba-1 Microglia Marker in Mouse Spinal Cord
Immunohistochemical staining of mouse spinal cord sections (L3–L5) from SOD1-G93A mice were double-immunostained either with anti-Iba-1, NeuN or GFAP (green) and Anti-P2X7 Receptor-ATTO-550 Antibody (#APR-004-AO), (red). P2X7 is present only in microglia cells as shown in the merged panel (yellow). Scale bar = 20 μm, insets = 50 μm.Adapted from Apolloni, S.et al. (2013) with permission of Oxford University Press.
Immuno-Colocalization of CaV1.2 and GABA(A) α1 Receptor in Rat Cerebellum
Immunohistochemical staining of rat cerebellum using Guinea pig Anti-CaV1.2 (CACNA1C) Antibody (#AGP-001) and Anti-GABA(A) α1 Receptor (extracellular) Antibody (#AGA-001). A. CaV1.2 (green) is detected in the granule layer of the cerebellum (G) and in the upper molecular layer (star). B. In the same section, GABA(A) α1 Receptor (red) is seen in the granule layer. C. Merge of the two images reveals high degree of colocalization between CaV1.2 and GABA(A) α1 Receptor in the granule layer.
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