BD, 556347, BD Pharmingen™ Purified Mouse Anti-VHL

Alternative Name: von Hippel-Lindau protein; RCA1; HRCA1
Reactivity: Human (QC Testing), Mouse (Reported)
Isotype: Mouse IgG1, κ
Immunogen: Human VHL aa. 1-213 Recombinant Protein
Application: Immunohistochemistry-formalin (antigen retrieval required) (Routinely Tested), Immunoprecipitation (Reported), Western blot (Not Recommended)
Target Molecular Weight: 21-30 kDa
Concentration: 0.5 mg/ml
RRID: AB_396376
Storage Buffer: Aqueous buffered solution containing ≤0.09% sodium azide.
Regulatory Status: RUO
Preparation And Storage: Preparation And Storage Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
Recommended Assay Procedures: Recommended Assay Procedures Western Blot : Clone Ig32 is not recommended for Western blot application.  It has been shown to recognize a non-specific band above VHL protein.  However, RNAi studies have demostrated that clone Ig32 recognizes a fainter band below this dominant band.  Also, labs have reported using the 786-O renal cell adenocarcinoma cell line (ATCC CRL-1932, VHL null), siRNA or VHL overexpression to show that clone Ig32 recognizes the VHL protein specifically.  Therefore, we recommend using clone Ig32 in over-expressing systems or under native (non-denaturing) conditions such as IHC staining or immunoprecipitation (IP) applications. Immunohistochemistry : The Ig32 antibody is useful for immunohistochemical staining. Following Retrievagen A pretreatment, purified Ig32 antibody should be used at 2.5 µg/ml to 5 µg/ml and titrated for optimal indirect immunohistochemical staining.  Tissues can be visualized via a three-step staining procedure in combination with Biotin Goat anti-Mouse Ig (Cat. No. 550337) secondary antibody and Streptravidin-HRP (Cat. No. 550946) together with the DAB Substrate Kit (Cat. No. 550880).  More conveniently, the Anti-Mouse Ig HRP Detection Kit (Cat. No. 551011) that contains the biotinylated secondary antibody, antibody diluent, streptavidin-HRP and DAB substrate can be used for staining.  For IHC staining on paraffin sections, please refer to http://www.bdbiosciences.com/documents/Immuno_Prep_Stain_Paraffin.pdf Additional protocol information can be found at http://www.bdbiosciences.com/support/resources/cell_biology/index.jsp IP : The purified Ig32 antibody has been reported to be useful to immunoprecipitate native human VHL.  Please note that this application is not routinely tested at BD Biosciences Pharmingen. Investigators are advised to determine optimal  concentrations for individual applications.