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Anti-TRPML3 (Mucolipin 3)-ATTO Fluor-594 Antibody

Mcoln3
Cat #: ACC-083-AR
Alternative Name Mcoln3
Lyophilized Powder yes
Type: Polyclonal
Host: Rabbit
Reactivity: h, m, r
Immunogen
  • Peptide CKDLPNSGKYRLEDD, corresponding to amino acid residues 528-542 of mouse TRPML3 (Accession Q8R4F0). Intracellular, C-terminus (cytoplasmic).
Accession (Uniprot) Number Q8R4F0
Gene ID 171166
Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
Homology Rat, human - identical.
RRID AB_10917153.
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.
Label ATTO-594. Maximum absorption 601 nm; maximum fluorescence 627 nm. The fluorescence is excited most efficiently in the 580 - 615 nm range. This label is related to the Rhodamine dyes and can be used with filters used to detect Texas Red and Alexa-594.
Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
Reconstitution 50 µl double distilled water (DDW).
Antibody concentration after reconstitution 1 mg/ml.
Storage after reconstitution The reconstituted solution can be stored at 4°C, protected from the light, 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 × g 5 min).
Standard quality control of each lot Western blot analysis (unlabeled antibody, #ACC-083), and immunohistochemistry (labeled antibody).
Applications: if, ih
May also work in: ic*
Immunohistochemistry
  • Expression of Mucolipin 3 in mouse brain
    Expression of Mucolipin 3 in mouse brain
    Immunohistochemical staining of TRPML3 in mouse brain frozen section with Anti-TRPML3 (Mucolipin 3)-ATTO Fluor-594 Antibody (#ACC-083-AR). A. Staining revealed TRPML3 positive nerve cell profiles in the brainstem auditory nucleus. B. Preincubation with the immunogen peptide suppressed the above staining pattern.
References
  1. Cheng, X. et al. (2010) FEBS Lett. 584, 2013.
  2. Christensen, K.A. et al. (2002) Curr. Opin. Cell. Biol. 17, 135.
  3. Lange, I. et al. (2009) Sci. Signal. 2, ra23.
  4. Calcraft, P.J. et al. (2009) Nature 459, 596.
  5. Puertollano, R. and Kiselyov, K. (2009) Am. J. Physiol. 296, F1245.
  6. Sun, M. et al. (2000) Hum. Mol. Genet. 9, 2471.
  7. Samie, M.A. et al. (2009) Pflugers Arch. 459, 79.
  8. Cuajungco, M.P. et al. (2008) Pflugers Arch. 457, 463.
  9. Nagata, K. et al. (2008) Proc. Natl. Acad. Sci. U.S.A. 105, 353.
  10. Kim, H.J. et al. (2007) J. Biol. Chem. 282, 36138.
  11. Di Palma, F. et al. (2002) Proc. Natl. Acad. Sci. U.S.A. 99, 14994.
Scientific background

The endolysosome system takes part in important cellular functions such as membrane trafficking, protein transport, autophagy and signal transduction1. Endosomes result from endocytosis of the plasma membrane and lysosomes (which are derived from late endosomes) conatin mainly hydrolytic enzymes and generally have a low internal pH1. Like the endoplasmic reticulum (ER), endolysosomes also store Ca2+ (luminal Ca2+ concentration: 0.5 mM)1,2, and similarly to Ca2+ release from the ER, Ca2+ from endolysosomes may also play an important role in various signaling events. To date such candidates include members of the TRP super-family of ion channels and the two-pore Ca2+ channels (TPCs)1,3,4.

TRPMLs, also termed mucolipins, are members of the TRP channels. In mammals, three TRPMLs are known to date (TRPML1-3 or MCOLN1-3). They are all localized to endolysosomes, although when over expressed in heterologous systems, TRPML3 is found on the plasma membrane1,5. These channels are Ca2+ permeable and display inward rectifying current properties1,5. Like all members of this family, TRPMLs have six transmembrane domains and intracellular N- and C-termini (relatively short tails compared to other members). They are characterized by an exceptionally large extracellular (luminal) loop between transmembrane domains 1 and 2, and N-glycosylation sites are present in the first extracellular (luminal) loop5.

In mammals, TRPML1 is expressed in a ubiquitous manner and shows highest expression in the brain, kidney, spleen, liver and heart1,6. TRPML2 and TRPML3 are less widely expressed. Interestingly, in mouse, two splice variants exist for TRPML2. The shorter variant is more broadly expressed and is dominant over the longer variant in the thymus, spleen and kidney1,7. TRPML3 is highly detected in the thymus, lung, kidney, spleen and eye1,7,8, some epithelial cells1,9 and brain10.

Pathologies related to these channels include type IV mucolipidosis, a neurodegenetative disease characterized by retardation and retinal degeneration caused by a loss of function mutation in the gene encoding TRPML1. In contrast, a gain of function mutation in TRPML3, in mice, causes deafness, and pigmentation defects11.

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: 16/08/2020

Anti-TRPML3 (Mucolipin 3) Antibody (#ACC-083) is a highly specific antibody directed against an epitope of the mouse protein. The antibody can be used in western blot and immunohistochemistry applications. It has been designed to recognize TRPML3 from human, rat and mouse samples.

Anti-TRPML3 (Mucolipin 3)-ATTO Fluor-594 Antibody (#ACC-083-AR) is directly labeled with an ATTO-594 fluorescent dye. ATTO dyes are characterized by strong absorption (high extinction coefficient), high fluorescence quantum yield, and high photo-stability. The ATTO-594 fluorescent label belongs to the class of Rhodamine dyes and can be used with fluorescent equipment typically optimized to detect Texas Red and Alexa-594. Anti-TRPML3 (Mucolipin 3)-ATTO Fluor-594 Antibody has been tested in immunohistochemical applications and is specially suited to experiments requiring simultaneous labeling of different markers.

For research purposes only, not for human use

Applications

Specifications

Scientific Background

Citations

Citations
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