Bestrophin 1 Antibody (E6-6)

Images

 
Western Blot: Bestrophin 1 Antibody (E6-6) [NB300-164] - Detection of Bestrophin (68 kDa) from human RPE cell lysate.

Product Details

Summary
Reactivity Hu, Po, Ca, Rt(-), PmSpecies Glossary
Applications WB, ICC/IF, IHC-Fr, IHC-P, IP
Clone
E6-6
Clonality
Monoclonal
Host
Mouse
Conjugate
Unconjugated

Order Details

Bestrophin 1 Antibody (E6-6) Summary

Immunogen
Synthetic peptide conjugated to KLH corresponding to the C-terminus of human Bestrophin 1 (KDHMDPYWALENRDEAHS) [UniProt# O76090]
Specificity
Bestrophin (E6-6)
Predicted Species
Primate (100%). Backed by our 100% Guarantee.
Isotype
IgG1 Kappa
Clonality
Monoclonal
Host
Mouse
Gene
BEST1
Purity
Unpurified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot 1:1000
  • Immunocytochemistry/Immunofluorescence Assay dependent
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin
  • Immunoprecipitation
Application Notes
By Western blot, this antibody recognizes a band at ~68 kDa representing Bestrophin.** It is also useful for immunofluorescence. **Please see protocol for treatment of cell extract. Use in Immunohistochemistry-Paraffin reported in scientific literature (PMID 24345323)
Reviewed Applications
Read 1 Review rated 5
using
NB300-164 in the following applications:

Publications
Read Publications using
NB300-164 in the following applications:

Reactivity Notes

NB300-164 recognizes the human protein. The immunogen shares 88% homology to canine, 94% to porcine, and 100% to chimpanzee. This antibody does not detect the rat protein. Other species have not been tested. Canine reactivity reported in scientific literature (PMID: 27519691). Porcine reactivity reported in scientific literature (PMID: 28687758).

Packaging, Storage & Formulations

Storage
Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
Buffer
Ascites
Preservative
0.1% Sodium Azide
Purity
Unpurified

Alternate Names for Bestrophin 1 Antibody (E6-6)

  • ARB
  • Best disease
  • BEST
  • bestrophin 1
  • bestrophin-1
  • BMD
  • TU15B
  • vitelliform macular dystrophy 2
  • Vitelliform macular dystrophy protein 2
  • VMD2RP50

Background

Best macular dystrophy (BMD) or vitelliform macular dystrophy (VMD2), is an autosomal form of macular degeneration, characterized by a depressed light peak in the electrooculogram (EOG). It is inherited and has an early onset. Bestrophin is a 68 kDa basolateral plasma membrane protein encoded by the VMD2 gene. Bestrophin's function is still unknown, but data suggests that it is a chloride channel that plays a role in generating the altered EOG in Best disease patients. In addition, Bestrophin is a useful biochemical and histological marker of RPE (retinal pigment epithelial cells) cells.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Customers Who Viewed This Item Also Viewed...

MAB1419
Species: Hu, Rt
Applications: IHC, CyTOF-ready, ICC, ICFlow
NBP2-35213
Species: Hu
AF1513
Species: Mu
Applications: WB, Simple Western, Flow, IHC, IP, CyTOF-ready
NBP1-86713
Species: Hu
Applications: IHC, IHC-P
NBP1-30027
Species: Hu, Mu, Rt
Applications: WB, ICC/IF, IHC, IHC-P
NB120-7164
Species: Hu, Mu, Rt, Fi
Applications: ELISA, ICC/IF, IHC-Fr, MiAr
NB100-56505
Species: Hu, Rt
Applications: WB, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, IP, CyTOF-ready
NBP2-20383
Species: Hu, Mu
Applications: WB, ICC/IF, IHC, IHC-P
NBP1-91258
Species: Hu, Mu
Applications: WB, Simple Western, ICC/IF, IHC, IHC-P
NBP2-02623
Species: Hu, Rt, Ca
Applications: WB, Flow, ICC/IF, IHC, IHC-P
NBP2-34260
Species: Hu, Mu, Po, Bv, Rb
Applications: WB, Flow, ICC/IF, IHC-Fr, IHC-P, IP, PAGE
NB100-74413
Species: Hu, Mu, Rb
Applications: WB, ICC/IF, IHC, IHC-P, IP
6449-TEC/CF
Species: Hu
7270-IL/CF
Species: Hu
NB110-3638
Species: Hu
Applications: WB, ELISA, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, RI
NBP1-89131
Species: Hu
Applications: ICC/IF, IHC, IHC-P
NB300-164
Species: Hu, Po, Ca, Rt(-), Pm
Applications: WB, ICC/IF, IHC-Fr, IHC-P, IP

Publications for Bestrophin 1 Antibody (NB300-164)(24)

We have publications tested in 5 confirmed species: Human, Canine, Porcine, Porcine , Primate.

We have publications tested in 6 applications: ICC/IF, IHC-Fr, IHC-P, IP, WB, WB,ICC/IF.


Filter By Application
ICC/IF
(11)
IHC-Fr
(1)
IHC-P
(1)
IP
(1)
WB
(8)
WB,ICC/IF
(1)
All Applications
Filter By Species
Human
(14)
Canine
(3)
Porcine
(2)
Porcine
(1)
Primate
(1)
All Species
Showing Publications 1 - 10 of 24. Show All 24 Publications.
Publications using NB300-164 Applications Species
Klingeborn M, Dismuke WM, Skiba NP et al. Directional Exosome Proteomes Reflect Polarity-Specific Functions in Retinal Pigmented Epithelium Monolayers Sci Rep 2017 Jul 07 [PMID: 28687758] (ICC/IF, Porcine) ICC/IF Porcine
Shim SH, Kim G, Lee DR et al. Survival of Transplanted Human Embryonic Stem Cell-Derived Retinal Pigment Epithelial Cells in a Human Recipient for 22 Months. JAMA Ophthalmol. 2017 Feb 09 [PMID: 28196199]
Kaneko Y, Pappas C, Tajiri N, Borlongan CV. Oxytocin modulates GABAAR subunits to confer neuroprotection in stroke in vitro. Sci Rep 2016 Oct 21 [PMID: 27767042]
Uggenti C, Briant K, Streit AK et al. Restoration of mutant bestrophin-1 expression, localisation and function. Dis Model Mech. Aug 12 2016 [PMID: 27519691] (WB,ICC/IF, Canine, Human) WB,ICC/IF Canine, Human
Parker J, Mitrousis N, Shoichet MS. Hydrogel for Simultaneous Tunable Growth Factor Delivery and Enhanced Viability of Encapsulated Cells in Vitro. Biomacromolecules. 2016 Feb 08 [PMID: 26762290] (ICC/IF, Human) ICC/IF Human
Bertolotti E, Neri A, Camparini M et al. Stem cells as source for retinal pigment epithelium transplantation. Prog Retin Eye Res. 2014 Jun 13 [PMID: 24933042] (ICC/IF, Human)

Details:
Figure 4. Basolateral localization of bestrophin 1 (antibody used 1:1000, Novus Biologicals) in human RPE cells. Human retinal stem cells (RNS; derived from adult ciliary bodies of post-mortem donors) were differentiated into RPE and the molecular characteristics of RPE cells were evaluated - polarization of the RPE cells was assessed with three markers: bestrophin 1 at the basolateral membrane, and beta5-integrin as well as ezrin at the apical side of the RPE.
ICC/IF Human
Sharon D, Al-Hamdani S, Engelsberg K et al. Ocular phenotype analysis of a family with biallelic mutations in the BEST1 gene. Am J Ophthalmol 2013 Dec 14 [PMID: 24345323] (IHC-P, ICC/IF, Human) IHC-P, ICC/IF Human
Maruotti J, Wahlin K, Gorrell D et al. A simple and scalable process for the differentiation of retinal pigment epithelium from human pluripotent stem cells. Stem Cells Transl Med 2013 May [PMID: 23585288]
Blenkinsop TA, Salero E, Stern JH, Temple S. The culture and maintenance of functional retinal pigment epithelial monolayers from adult human eye Methods Mol Biol 2013 [PMID: 23097100]
Cho MS, Kim SJ, Ku SY et al. Generation of retinal pigment epithelial cells from human embryonic stem cell-derived spherical neural masses Stem Cell Res 2012 May 16 [PMID: 22683799] (ICC/IF, Human) ICC/IF Human
Show All 24 Publications.

Review for Bestrophin 1 Antibody (NB300-164) (1) 51

Average Rating: 5
(Based on 1 review)
We have 1 review tested in 1 species: Human and Rat.
We have 1 review tested in 1 application: WB.

Reviews using NB300-164:
Filter by Applications
WB
(1)
All Applications
Filter by Species
Human and Rat
(1)
All Species
Images Ratings Applications Species Date Details
Western Blot Bestrophin 1 NB300-164
Enlarge
reviewed by:
Anonymous
WB Human and Rat 12/29/2016
View

Summary

ApplicationWestern Blot
Sample TestedRPE whole cell lysate
SpeciesHuman and Rat

Comments

CommentsE6-6 used at 1:2000 over night. next day secondary antibody donkey anti-mouse peroxidase was applied followed by detection using enhanced chemiluminescence.

Product General Protocols

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for Bestrophin 1 Antibody (NB300-164). (Showing 1 - 4 of 4 FAQs).

  1. According to the data sheet, for Immunohistochemistry, it references (PMID: 21249129). I see a 1:25 dilution in the journal article referenced under immunocytochemistry. Is this the same dilution recommended for IHC? If not do you have a recommended dilution? Antigen retrieval: Do you have any information on AR? Citrate buffer or higher pH (EDTA)? Temperature? Time? Incubation: Is 1 hour at room temperature or overnight at 4C recommended?
    • This antibody was validated in IHC in the reference PMID 11050159. Generally we recommend using antigen retrieval with citrate buffer and incubating for 1 hour at RT. Some antigens may be tricky and require overnight incubation. I imagine the specific protocol can be found in the reference, however.
  2. Can I get the recipe for RIPA buffer and the lysate preparation protocol for membrane bound proteins using NB300-164?
    • Here is both the recipe we use for RIPA and a good protocol for tissue lysate prep. RIPA buffer: 150 mM sodium chloride 1.0% NP-40 or Triton X-100 0.5% sodium deoxycholate 0.1% SDS (sodium dodecyl sulphate) 50 mM Tris, pH 8.0 RIPA buffer is also useful for whole cell extracts and membrane-bound proteins, and may be preferable to NP-40 or Triton X100-only buffers for extracting nuclear proteins. It will disrupt protein-protein interactions and may therefore be problematic for immunoprecipitations/pull down assays. Preparation of lysate from tissues Dissect the tissue of interest with clean tools, on ice preferably, and as quickly as possible to prevent degradation by proteases. Place the tissue in round-bottom microfuge tubes or Eppendorf tubes and immerse in liquid nitrogen to snap freeze. Store samples at -80C for later use or keep on ice for immediate homogenization. For a approximately 5 mg piece of tissue, add approximately 300 ul lysis buffer rapidly to the tube, homogenize with an electric homogenizer, rinse the blade twice with another 2x300 ul lysis buffer, then maintain constant agitation for 2 hours at 4C (e.g place on an orbital shaker in the fridge). Volumes of lysis buffer must be determined in relation to the amount of tissue present (protein extract should not be too diluted to avoid loss of protein and large volumes of samples to be loaded onto gels. The minimum concentration is 0.1 mg/ml, optimal concentration is 1-5 mg/ml). Centrifuge for 20 min at 12000 rpm at 4C in a microcentrifuge. Gently remove the tubes from the centrifuge and place on ice, aspirate the supernatant and place in a fresh tube kept on ice; discard the pellet.
  3. I want to know the concentration of Bestrophin 1 antibody [NB300-164].
    • For our product NB300-164, this antibody is provided as unpurified ascites. Therefore, the antibody concentration has not been determined.
  4. We are interested in your item, NB300-164 and would you please let us know the antibody concentration of it?
    • The total protein concentration of the ascites (Lot# B-3) is 11.7 mg/ml.

Secondary Antibodies

 

Isotype Controls

Additional Bestrophin 1 Products

Bioinformatics Tool for Bestrophin 1 Antibody (NB300-164)

Discover related pathways, diseases and genes to Bestrophin 1 Antibody (NB300-164). Need help? Read the Bioinformatics Tool Guide for instructions on using this tool.
Visit Tool

Diseases for Bestrophin 1 Antibody (NB300-164)

Discover more about diseases related to Bestrophin 1 Antibody (NB300-164).
 

Pathways for Bestrophin 1 Antibody (NB300-164)

View related products by pathway.

PTMs for Bestrophin 1 Antibody (NB300-164)

Learn more about PTMs related to Bestrophin 1 Antibody (NB300-164).
 

Research Areas for Bestrophin 1 Antibody (NB300-164)

Find related products by research area.

Blogs on Bestrophin 1.

Vision Infographic: Do you see how I see?
Vision involves several parts of the eye processing light which send signals to the brain via the optic nerve to process information. Learn more about the vision process and related ocular proteins in the infographic below. Novus Biological...  Read full blog post.

Bestrophin 1: Implications in Progressive Vision Loss
The human Bestrophin family has four members, Best1, Best2, Best3 and Best4. These transmembrane proteins can function as chloride channels, and can also regulate calcium channels (1). The Bestrophins all have a conserved domain which begins at the N-...  Read full blog post.

"I can see clearly now": Targeting Bestrophin 1 to treat Bestrophinopathies
Encoded by the VMD2 gene on chromosome 11q13 Bestrophin 1 is the prototypic member of the RFP family of proteins which are more commonly called "bestrophins". The protein family was originally identified in Caenorhabditis elegans based on a conserved...  Read full blog post.

Customers Who Bought This Also Bought

Contact Information

Product PDFs

Calculators

Concentration Calculator

The concentration calculator allows you to quickly calculate the volume, mass or concentration of your vial. Simply enter your mass, volume, or concentration values for your reagent and the calculator will determine the rest.

=
÷

Recent Reviews

5
5
1
4
0
3
0
2
0
1
0

Anonymous
12/29/2016
Application: WB
Species: Human and Rat

Bioinformatics

Gene Symbol BEST1
Entrez
Uniprot