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Anti-NETO2 (extracellular) Antibody

Neuropilin and tolloid-like protein 2, Brain-specific transmembrane protein containing 2 CUB and 1 LDL-receptor class A domains protein 2, BTCL2
Cat #: AGC-046
Alternative Name Neuropilin and tolloid-like protein 2, Brain-specific transmembrane protein containing 2 CUB and 1 LDL-receptor class A domains protein 2, BTCL2
Lyophilized Powder yes
Type: Polyclonal
Host: Rabbit
Reactivity: h, m, r
Immunogen
  • Peptide (C)RSSQVEQEEKTKPG, corresponding to amino acid residues 189-202 of rat NETO2 (Accession C6K2K4). Extracellular, N-terminus.
Accession (Uniprot) Number C6K2K4
Gene ID 307757
Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
Homology Mouse, human – identical.
Purity Affinity purified on immobilized antigen.
Form Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.05% NaN3.
Isotype Rabbit IgG.
Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
Reconstitution 25 µl, 50 µl or 0.2 ml double distilled water (DDW), depending on the sample size.
Antibody concentration after reconstitution 0.8 mg/ml.
Storage after reconstitution The reconstituted solution can be stored at 4°C for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 x g 5 min).
Standard quality control of each lot Western blot analysis.
Applications: if, ih, wb
May also work in: ic*, ifc*, ip*, lci*
Western blot
  • Western blot analysis of mouse (lanes 1 and 3) and rat (lanes 2 and 4) brain lysates:
    Western blot analysis of mouse (lanes 1 and 3) and rat (lanes 2 and 4) brain lysates:
    1,2. Anti-NETO2 (extracellular) Antibody (#AGC-046), (1:200).
    3,4. Anti-NETO2 (extracellular) Antibody, preincubated with NETO2 (extracellular) Blocking Peptide (#BLP-GC046).
  • Western blot analysis of human SH-SY5Y neuroblastoma (lanes 1 and 4), human HT-29 colon adenocarcinoma (lanes 2 and 5) and human MCF-7 breast adenocarcinoma (lanes 3 and 6) cell line lysates:
    Western blot analysis of human SH-SY5Y neuroblastoma (lanes 1 and 4), human HT-29 colon adenocarcinoma (lanes 2 and 5) and human MCF-7 breast adenocarcinoma (lanes 3 and 6) cell line lysates:
    1-3. Anti-NETO2 (extracellular) Antibody (#AGC-046), (1:200).
    4-6. Anti-NETO2 (extracellular) Antibody, preincubated with NETO2 (extracellular) Blocking Peptide (#BLP-GC046).
Immunohistochemistry
  • Expression of NETO2 in mouse and rat cerebellum
    Expression of NETO2 in mouse and rat cerebellum
    Immunohistochemical staining of perfusion-fixed frozen mouse and rat brain sections with Anti-NETO2 (extracellular) Antibody (#AGC-046), (1:200), followed by goat anti-rabbit-AlexaFluor-488. NETO2 immunoreactivity (green) in both mouse (A.) and rat (B.) cerebellum region is detected in the granule layer (G) and in “baskets” (arrows). Cell nuclei are stained with DAPI (blue).
References
  1. Fisher, J.L. (2015) Neuropharmacology 99, 471.
  2. Fisher, J.L. and Mott, D.D. (2013) J. Physiol. 591, 4711.
  3. Lomash, R.M. et al. (2017) J. Biol. Chem. 292, 15369.
Scientific background

NETO proteins are auxiliary subunits that play a crucial role in the modulation of the biophysical properties of Kainate-type glutamate receptors (KARs). KARs are activated to modulate synaptic transmission, network excitability and synaptogenesis. NETO proteins modulate their activity both in vivo and in vitro mostly by regulating the surface expression and synaptic targeting of KARs in neurons.

NETO protein expression has been characterized in the adult brain. NETO1 and NETO2 are highly homologous but have different expression patterns and distinct functional effects. High NETO2 expression levels can be found throughout the mammalian brain1,2.

The NETO2 structure reveals a single-pass transmembrane protein that contains an extracellular region with two CUB domains that mediates interactions with KAR subunits. The C-terminus is important for modifying and regulating the synaptic abundance of KARs. The intracellular region of NETO2 can be phosphorylated by various kinases in vitro,which regulates synaptic targeting of KARs3.

Application key:

CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

Species reactivity key:

H- Human, M- Mouse, R- Rat
Last update: 23/08/2020

Anti-NETO2 (extracellular) Antibody (#AGC-046) is a highly specific antibody directed against an epitope of the rat protein. The antibody can be used in western blot and immunohistochemistry applications. It has been designed to recognize NETO2 from human, rat, and mouse samples.

For research purposes only, not for human use
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