VHL antibody

Catalog No. ABIN2689963

Product Details anti-VHL Antibody

Target: VHL (Von Hippel-Lindau Tumor Suppressor, E3 Ubiquitin Protein Ligase (VHL))

Reactivity: Human

Host: Mouse

Clonality: Monoclonal

Conjugate: This VHL antibody is un-conjugated

Application: Western Blotting (WB), Immunohistochemistry (Formalin-fixed Sections) (IHC (f)), Intracellular Staining (ICS)

Brand: BD Pharmingen™

Characteristics: The S2-647 monoclonal antibody recognizes the Von Hippel-Lindau (VHL) protein. The VHL protein is expressed within the cytosol and nuclei of cells from a variety of tissues. It serves as a tumor suppressor and transcriptional repressor that is highly conserved from Drosophila to mammalian species. VHL binds to Elongin B and C subunits and other proteins as part of a E3 ubiquitin ligase complex that targets a variety of proteins, including HIF-1 alpha and Beta-2 adrenergic receptors, for proteasomal degradation. Aberrant VHL expression is associated with a number of diseases including Von Hippel-Lindau disease, a familial cancer syndrome, that predisposes individuals to hemangioblastomas of the central nervous system and retina, pheochromocytoma and renal cell carcinoma. Three distinct VHL isoforms ranging in molecular weight from approximately 19-30 kDa have been described. Western blot analysis of VHL expression and RNAi validation of anti-VHL antibody specificity - Panel 1. Lysate prepared from HeLa S3 cells (Panel 1a) was blotted using Purified Mouse Anti-VHL antibody (Cat. No. 564183) at concentrations of 1 (lane 1), 0.5 (lane 2), or 0.25 (lane 3) μg/mL. VHL was identified as a band of ~24 kDa (Panel 1a). Lysates from untreated, VHL RNAi- or ERK2 RNAi-treated HeLa S3 cells (Panel 1b) were blotted with Purified Mouse Anti-VHL or Anti-ERK-2 (Cat. No. 554095) antibody. The expressed level of VHL protein blotted with Anti-VHL antibody (2 μg/mL) is downregulated in lysate prepared from HeLa S3 cells treated with VHL RNAi but not ERK2 RNAi. Similarly, ERK2 protein expression was reduced in lysate from HeLa S3 cells treated with ERK2 RNAi but not VHL RNAi when blotted with Anti-ERK2 antibody (0.5 μg/mL). Immunohistochemical staining of VHL expressed in tissue from a human renal tumor - Panel 2. Following antigen retrieval with BD Retrievagen A buffer (Cat. no. 550524), the formalin-fixed paraffin-embedded sections were stained with either Purified Mouse IgG2b, κ Isotype Control (Cat. No. 557351, Top Tissue) or Purified Mouse Anti-VHL antibody (Bottom Tissue) using the BD Biosciences Protocol for Immunohistochemical Staining. A three-step staining procedure that employs a Biotin Goat Anti-Mouse Immunoglobulin (Cat. No. 550337), Streptavidin-Horseradish Peroxidase (HRP) (Cat. No.550946), and the DAB Substrate Kit (Cat. No. 550880) was used to develop the primary staining reagents. The tissue was counterstained with Hematoxylin. Original magnification: 20X. Flow cytometric analysis of VHL expression in VHL-transfected 293F cells - Panel 3. Untransfected (dashed line histogram) and human VHL-transfected (solid line histogram) 293F cells were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were then washed and stained with Purified Mouse Anti-Human VHL antibody followed by APC Goat Anti-Mouse Ig (Cat. No. 550826) using BD Biosciences Protocol for Intracellular Staining. The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometric analysis was performed using a BD FACSCanto™ II Flow Cytometer System. 564183 Rev. 1 Page 1 of 2

BD Pharmingen™ Purified Mouse Anti-Human VHL - Purified - Clone S2-647 - Isotype Mouse IgG2b, κ - Reactivity Hu - 0.1 mg

Purification: The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Immunogen: Human VHL Recombinant Protein

Clone: S2

Isotype: IgG2b kappa

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Restrictions: For Research Use only

Handling

Concentration: 0.5 mg/mL

Buffer: Aqueous buffered solution containing ≤0.09 % sodium azide.

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: 4 °C

Storage Comment: Store undiluted at 4°C.

References for anti-VHL antibody (ABIN2689963)

Qi, Gervais, Li, DeCaprio, Challis, Ohh: "Molecular cloning and characterization of the von Hippel-Lindau-like protein." in: Molecular cancer research : MCR, Vol. 2, Issue 1, pp. 43-52, (2004) (PubMed).

Baba, Hirai, Kawakami, Kishida, Sakai, Kaneko, Yao, Shuin, Kubota, Hosaka, Ohno: "Tumor suppressor protein VHL is induced at high cell density and mediates contact inhibition of cell growth." in: Oncogene, Vol. 20, Issue 22, pp. 2727-36, (2001) (PubMed).

Iliopoulos, Ohh, Kaelin: "pVHL19 is a biologically active product of the von Hippel-Lindau gene arising from internal translation initiation." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 95, Issue 20, pp. 11661-6, (1998) (PubMed).

Chen, Kishida, Duh, Renbaum, Orcutt, Schmidt, Zbar: "Suppression of growth of renal carcinoma cells by the von Hippel-Lindau tumor suppressor gene." in: Cancer research, Vol. 55, Issue 21, pp. 4804-7, (1995) (PubMed).

Iliopoulos, Kibel, Gray, Kaelin: "Tumour suppression by the human von Hippel-Lindau gene product." in: Nature medicine, Vol. 1, Issue 8, pp. 822-6, (1995) (PubMed).

Kibel, Iliopoulos, DeCaprio, Kaelin: "Binding of the von Hippel-Lindau tumor suppressor protein to Elongin B and C." in: Science (New York, N.Y.), Vol. 269, Issue 5229, pp. 1444-6, (1995) (PubMed).

Target Details for VHL

Target: VHL (Von Hippel-Lindau Tumor Suppressor, E3 Ubiquitin Protein Ligase (VHL))

Alternative Name: VHL (VHL Products)

Synonyms: HRCA1 antibody, RCA1 antibody, VHL1 antibody, pVHL antibody, BcDNA:RH61560 antibody, CG13221 antibody, DVhl antibody, Dmel\\CG13221 antibody, Dvhl antibody, VHL antibody, d-VHL antibody, d-vhl antibody, dVHL antibody, dmVHL antibody, vhl antibody, rca1 antibody, vhl1 antibody, hrca1 antibody, zgc:158722 antibody, Vhlh antibody, von Hippel-Lindau tumor suppressor antibody, von Hippel-Lindau antibody, von Hippel-Lindau tumor suppressor, E3 ubiquitin protein ligase antibody, von Hippel-Lindau disease tumor suppressor antibody, VHL antibody, Vhl antibody, vhl antibody, CpipJ_CPIJ009992 antibody

Background: Synonyms: VHL1, pVHL, Elongin binding protein, Protein G7, HRCA1, RCA1

Molecular Weight: 23-25 kDa

Pathways: Tube Formation, Signaling Events mediated by VEGFR1 and VEGFR2, Ubiquitin Proteasome Pathway