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Anti-Bestrophin-1 (extracellular) Antibody

BEST1, Vitelliform macular dystrophy protein 2, VMD2, BMD1

Cat #: ABC-001
Alternative Name BEST1, Vitelliform macular dystrophy protein 2, VMD2, BMD1
Lyophilized Powder yes
Type: Polyclonal
Host: Rabbit
Reactivity: h, m, r
Immunogen
  • Peptide (C)NPNKDYPGHEMD, corresponding to amino acid residues 259-270 of mouse Bestrophin-1 (Accession O88870). 3rd extracellular loop.
Accession (Uniprot) Number O88870
Gene ID 24115
Peptide confirmation Confirmed by amino acid analysis and mass spectrometry.
Homology Rat - 10/12 amino acid residues identical; human - 9/12 amino acid residues identical.
RRID AB_10560217.
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: ifc, ih, wb
May also work in: ic*, ip*, lci*
Western blot
  • Western blot analysis of rat lung (lanes 1 and 3) and rat eye (lanes 2 and 4) lysate:
    Western blot analysis of rat lung (lanes 1 and 3) and rat eye (lanes 2 and 4) lysate:
    1,2. Anti-Bestrophin-1 (extracellular) Antibody (#ABC-001), (1:200).
    3,4. Anti-Bestrophin-1 (extracellular) Antibody, preincubated with Bestrophin-1 (extracellular) Blocking Peptide (#BLP-BC001).
Immunohistochemistry
  • Expression of Bestrophin-1 in rat lung
    Expression of Bestrophin-1 in rat lung
    Immunohistochemical staining of paraffin embedded rat lung sections using Anti-Bestrophin-1 (extracellular) Antibody (#ABC-001), (1:100). BEST1 is expressed both in the respiratory epithelium (red arrows) and in vascular smooth muscle (black arrows). Hematoxilin is used as the counterstain.
Indirect flow cytometry
  • Cell surface detection of Bestrophin-1 in live intact Jurkat (human T cell leukemia) cell line:
    Cell surface detection of Bestrophin-1 in live intact Jurkat (human T cell leukemia) cell line:
    ___ Control cells + goat-anti-rabbit-FITC.
    ___ Cells + Anti-Bestrophin-1 (extracellular) Antibody (#ABC-001)(1:20) + goat-anti-rabbit-FITC.
  • The control antigen is not suitable for this application.
References
  1. Men, G. et al. (2004) Am. J. Ophtalmol. 137, 963.
  2. Petrukhin, K. et al. (1998) Nat. Genet. 19, 241.
  3. Hartzell, C.H. et al. (2008) Physiol. Rev. 88, 639.
  4. Kramer, F. et al. (2004) Cytogen. Genome Res. 105, 107.
  5. Stohr, H. et al. (2002) Eur. J. Hum. Genet. 10, 281.
  6. Park, H. et al. (2009) J. Neurosci. 29, 13063.
  7. Marmostein, A.D. et al. (2009) Prog Retin Eye Res. 28, 206.
  8. Tsunenari, T. et al. (2003) J. Biol. Chem. 278, 41114.
  9. Milenkovic, V.M. et al. (2007) J. Biol. Chem. 282, 1313.
  10. Xiao, Q. et al. (2008) J. Gen. Physiol. 132, 681.
  11. Chien, L.T. et al. (2006) J. Gen. Physiol. 128, 247.
  12. Tsunenari, T. et al. (2006) J. Gen. Physiol. 127, 749.
  13. Woo, D.H. et al. (2012) Cell 151, 25.
Scientific background

Mammalian Cl- channels can be broadly classified into four different families: voltage-dependent Cl- channels (CLCs), the cystic fibrosis transmembrane conductance regulator (CFTR), ligand-gated Cl- channels (γ-aminobutyric acid (GABA)) and glycine channels) and Ca2+-activated Cl- channels (Bestrophin and Anoctamin channels).

Bestrophins were first found by genetic linkage of human-Bestrophin-1 (hBest1) to a juvenile form of macular degeneration called Best vitelliform macular dystrophy (BVMD)1,2. BVMD is mainly electrophysiologically characterized by a decrease in the light peak and physiologically by the thinning of the retina layer which eventually leads to the loss of central vision3. To date Bestrophin 1-4 have been identified, although Bestrophin-3 and Bestrophin-4 have been observed only at the RNA level3. In addition, splice variants of some of these Ca2+-activated Cl- channels (CaCCs) have also been detected2,4,5. CaCCs are known to be involved in the regulation of olfaction, taste, phototransduction, and excitability in the nervous system. Recently, Bestrophin-1 was shown to be functionally expressed in astrocytes in both primary cell culture and in situ6. Bestrophin-1 is also detected in retina, brain, spinal cord and testes7.

Two different topologies for Bestrophin-1 have been proposed. The first, the preferred structure, proposes that six hydrophobic domains span the membrane8, while the second suggests that there are only four membrane-spanning domains9. Bestrophin-1, along with its counterparts, is activated by intracellular Ca2+. A recent study demonstrated that Bestrophin-1 indeed binds Ca2+ and by mutating specific residues, showed which amino acid residues are essential for binding Ca2+ 10, providing additional evidence that Bestrophin-1 is activated by direct binding of Ca2+ to the channel11,12.

Bestrophin-1 has been found to release glutamate from astrocytes and is located at microdomains near synapses13.

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: 11/04/2021

Alomone Labs is pleased to offer a highly specific antibody directed against an extracellular epitope of mouse Bestrophin-1. Anti-Bestrophin-1 (extracellular) Antibody (#ABC-001) can be used in western blot, immunohistochemistry and indirect flow cytometry applications. It has been designed to recognize bestrophin-1 channel from mouse, rat and human samples.

For research purposes only, not for human use

Applications

Specifications

Scientific Background

Citations

Citations
Western blot citations
  1. Rat uterus lysate (1:2000).
    Mijuškovic´, A. et al. (2015) Br. J. Pharmacol. 172, 3671.
  2. Rat spinal cord and DRG lysates (1:200).
    Pineda-Farias, J.B. et al. (2015) Mol. Pain 11, 1.
  3. Rat DRG lysate (1:400).
    Garcia, G. et al. (2014) Brain Res. 1579, 35.
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