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)
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.
Alternate Names for Bestrophin 1 Antibody (E6-6)
vitelliform macular dystrophy 2
Vitelliform macular dystrophy protein 2
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.
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.
FAQs for Bestrophin 1 Antibody (NB300-164). (Showing 1 - 4 of 4 FAQs).
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.
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.
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.
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.
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