Invitrogen

VHL Polyclonal Antibody

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3 Published Figures

View all (22) VHL antibodies

Questions & Answers

Questions & Answers

VHL Antibody in Immunocytochemistry (ICC/IF)

VHL Antibody in Immunocytochemistry (ICC/IF)

FIGURE: 1 / 9

VHL Antibody (PA5-27322) in ICC/IF

Immunocytochemistry-Immunofluorescence analysis of VHL was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: VHL Polyclonal Antibody (Product # PA5-27322) diluted at 1:500. Red: phalloidin, a cytoskeleton marker. Scale bar = 10 µm.

VHL Antibody in Immunocytochemistry (ICC/IF)

VHL Antibody in Immunohistochemistry (Paraffin) (IHC (P))

VHL Antibody in Immunohistochemistry (Paraffin) (IHC (P))

VHL Antibody in Western Blot (WB)

VHL Antibody in Western Blot (WB)

VHL Antibody in Western Blot, Immunocytochemistry (WB, ICC/IF)

VHL Antibody in Western Blot (WB)

VHL Antibody in Immunoprecipitation (IP)

Product Image Gallery

VHL Antibody in Immunocytochemistry (ICC/IF) — Immunocytochemistry-Immunofluorescence analysis of VHL was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: VHL Polyclonal Antibody (Product # PA5-27322) diluted at 1:500. Red: phalloidin, a cytoskeleton marker. Scale bar = 10 µm.

VHL Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry (Paraffin) analysis of VHL was performed in paraffin-embedded mouse kidney tissue using VHL Polyclonal Antibody (Product # PA5-27322) at a dilution of 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

VHL Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry (Paraffin) analysis of VHL was performed in paraffin-embedded mouse lung tissue using VHL Polyclonal Antibody (Product # PA5-27322) at a dilution of 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

VHL Antibody in Western Blot (WB) — Western Blot analysis of VHL was performed by separating 30 µg of non-transfected (–) and transfected (+) 293T whole cell extracts by 12% SDS-PAGE. Proteins were transferred to a membrane and probed with a VHL Polyclonal Antibody (Product # PA5-27322) at a dilution of 1:5000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

VHL Antibody in Western Blot (WB) — Western Blot using VHL Polyclonal Antibody (Product # PA5-27322). Various whole cell extracts (30 µg) were separated by 12% SDS-PAGE, and the membrane was blotted with VHL Polyclonal Antibody (Product # PA5-27322) diluted at 1:500. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

VHL Antibody in Western Blot, Immunocytochemistry (WB, ICC/IF) — Published figure using VHL polyclonal antibody (Product # PA5-27322) in Western Blot and Immunocytochemistry

VHL Antibody in Western Blot (WB) — Figure 3 miR-147b-VHL axis mediates drug-tolerance through impaired VHL activity. a , Left, gene candidates predicted for miR-147b by the TargetScan tool were shown in signaling pathways enriched for gefitinib-tolerance in PC9 single-cell clones in fig. 1f . Right, qRT-PCR analysis for the predicted gene candidates for miR-147b in H1975 cells with miR-147b knockdown compared with scrambled control. n=3 independent biological replicates. b , Left, computational prediction of RNA duplex formation between miR-147b and the 3'UTR (untranslated region) of VHL mRNA. Mutations generated within the 3'UTR for the luciferase assay are shown in red. Right, dual-luciferase reporter assay in miR-147b-overexpressing AALE cells. The Firefly luciferase and Renilla luciferase activities were measured 48 hours post co-transfection with miR-147b or control vector and wild-type (WT) or mutant (Mut) VHL 3'UTR. n=3 independent biological replicates. c , Western blot analysis and quantification of VHL in miR-147b-overexpressing AALE cells. beta-Actin was used as loading control. n=3 independent biological replicates. d , qRT-PCR analysis for fold change of hypoxia gene expression in AALE cells with miR-147b overexpression relative to scrambled control (147b/Scr) and cells with co-overexpression of miR-147b and VHL relative to scrambled control (147b+VHL/Scr). ACTB was used as endogenous control. n=3 independent biological replicates. e , Fractional viability of HCC827 cells treated with vehicle, osi

VHL Antibody in Immunoprecipitation (IP) — Published figure using VHL polyclonal antibody (Product # PA5-27322) in Immunoprecipitation

VHL Antibody — Figure 3. UBE2C mediates shear-dependent expression of pVHL and HIF-1&agr; in human aortic valve endothelial cells (HAVECs), and UBE2C, pVHL, and HIF1&agr; are expressed in a side-dependent manner in human aortic valve (AV) leaflets. A , Depicts a hypothesis that overexpression of UBE2C ubiquitinates pVHL, leading to increased HIF-1&agr; level, endothelial inflammation, and endothelial-mesenchymal-transition (EndMT). B , HAVECs sheared for 72 h by stable flow (SF) or disturbed flow (DF) were immunostained with antibodies for pVHL or HIF-1&agr; ( B ) and quantification of the image was done using ImageJ ( C ), or by qPCR ( D ) normalized to 18S (n=3-4). E , HAVECs treated with siCtrl or siUBE2C for 24 h, followed by exposure to DF for 24 h, were analyzed by Western blot ( E ) and Image J quantification ( F ) for pVHL (n=5). For the HIF-1&agr; study, HAVECs exposed to DF for 72 h were immunostained with HIF-1&agr; ( G ) and Image J quantified ( H ; n=6). I , Human AVs with sclerosis were stained with antibodies to UBE2C, pVHL, HIF-1&agr;, Twist1, and Runx2 (runt-related transcription factor 2) with DAPI nuclear staining and ( J ) show quantification of the fluorescent intensities of each staining in the endothelial layer on each side using Image J (n=5-8). Mean +- SEM, * P <0.05. Fib indicates fibrosa; and Ven, ventricularis. {TM}

Product Details

PA5-27322

Applications

Tested Dilution

Publications

Western Blot (WB)

1:500-1:10,000

View 2 publications 2 publications

Immunohistochemistry (Paraffin) (IHC (P))

1:100-1:1,000

-

Immunocytochemistry (ICC/IF)

1:100-1:1,000

View 1 publication 1 publication

Immunoprecipitation (IP)

Assay-dependent

View 1 publication 1 publication

Product Specifications

Species Reactivity

Human, Mouse

Published species

Human

Host/Isotype

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

Recombinant protein encompassing a sequence within the center region of human Von Hippel Lindau. The exact sequence is proprietary.

Conjugate

Unconjugated Unconjugated Unconjugated

Form

Liquid

Concentration

0.6 mg/mL

Purification

Antigen affinity chromatography

Storage buffer

PBS, pH 7, with 20% glycerol

Contains

0.025% ProClin 300

Storage conditions

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Shipping conditions

Ambient (domestic); Wet ice (international)

AB_2544798

Product Specific Information

Recommended positive controls: Myc-DDK-tagged VHL-transfected 293T.

Predicted reactivity: Mouse (93%), Rat (91%), Dog (95%), Bovine (96%).

Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Target Information

Von Hippel-Lindau syndrome (VHL) is a dominantly inherited familial cancer syndrome predisposing to a variety of malignant and benign tumors. A germline mutation of this gene is the basis of familial inheritance of VHL syndrome. The protein encoded by this gene is a component of the protein complex that includes elongin B, elongin C, and cullin-2, and possesses ubiquitin ligase E3 activity. This protein is involved in the ubiquitination and degradation of hypoxia-inducible-factor (HIF), which is a transcription factor that plays a central role in the regulation of gene expression by oxygen. RNA polymerase II subunit POLR2G/RPB7 is also reported to be a target of this protein. Alternatively spliced transcript variants encoding distinct isoforms have been observed.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

Disturbed Flow Increases UBE2C (Ubiquitin E2 Ligase C) via Loss of miR-483-3p, Inducing Aortic Valve Calcification by the pVHL (von Hippel-Lindau Protein) and HIF-1α (Hypoxia-Inducible Factor-1α) Pathway in Endothelial Cells.

Arteriosclerosis, thrombosis, and vascular biology

Fernandez Esmerats J,Villa-Roel N,Kumar S,Gu L,Salim MT,Ohh M,Taylor WR,Nerem RM,Yoganathan AP,Jo H

PA5-27322 was used in Western Blotting to test the role of novel flow-sensitive UBE2C (ubiquitin E2 ligase C) and microRNA-483-3p (miR-483) in flow-dependent aortic valve (AV) endothelial function and AV calcification.

Fri Mar 01 00:00:00 UTC 2019

Disturbed Flow Increases UBE2C (Ubiquitin E2 Ligase C) via Loss of miR-483-3p, Inducing Aortic Valve Calcification by the pVHL (von Hippel-Lindau Protein) and HIF-1α (Hypoxia-Inducible Factor-1α) Pathway in Endothelial Cells.

Arteriosclerosis, thrombosis, and vascular biology

Fernandez Esmerats J,Villa-Roel N,Kumar S,Gu L,Salim MT,Ohh M,Taylor WR,Nerem RM,Yoganathan AP,Jo H

PA5-27322 was used in Western Blotting to test the role of novel flow-sensitive UBE2C (ubiquitin E2 ligase C) and microRNA-483-3p (miR-483) in flow-dependent aortic valve (AV) endothelial function and AV calcification.

Fri Mar 01 00:00:00 UTC 2019

Disturbed Flow Increases UBE2C (Ubiquitin E2 Ligase C) via Loss of miR-483-3p, Inducing Aortic Valve Calcification by the pVHL (von Hippel-Lindau Protein) and HIF-1α (Hypoxia-Inducible Factor-1α) Pathway in Endothelial Cells.

Arteriosclerosis, thrombosis, and vascular biology

Fernandez Esmerats J,Villa-Roel N,Kumar S,Gu L,Salim MT,Ohh M,Taylor WR,Nerem RM,Yoganathan AP,Jo H

PA5-27322 was used in Western Blotting to test the role of novel flow-sensitive UBE2C (ubiquitin E2 ligase C) and microRNA-483-3p (miR-483) in flow-dependent aortic valve (AV) endothelial function and AV calcification.

Fri Mar 01 00:00:00 UTC 2019

Disturbed Flow Increases UBE2C (Ubiquitin E2 Ligase C) via Loss of miR-483-3p, Inducing Aortic Valve Calcification by the pVHL (von Hippel-Lindau Protein) and HIF-1α (Hypoxia-Inducible Factor-1α) Pathway in Endothelial Cells.

Arteriosclerosis, thrombosis, and vascular biology

Fernandez Esmerats J,Villa-Roel N,Kumar S,Gu L,Salim MT,Ohh M,Taylor WR,Nerem RM,Yoganathan AP,Jo H

PA5-27322 was used in Western Blotting to test the role of novel flow-sensitive UBE2C (ubiquitin E2 ligase C) and microRNA-483-3p (miR-483) in flow-dependent aortic valve (AV) endothelial function and AV calcification.

Fri Mar 01 00:00:00 UTC 2019

Disturbed Flow Increases UBE2C (Ubiquitin E2 Ligase C) via Loss of miR-483-3p, Inducing Aortic Valve Calcification by the pVHL (von Hippel-Lindau Protein) and HIF-1α (Hypoxia-Inducible Factor-1α) Pathway in Endothelial Cells.

Arteriosclerosis, thrombosis, and vascular biology

Fernandez Esmerats J,Villa-Roel N,Kumar S,Gu L,Salim MT,Ohh M,Taylor WR,Nerem RM,Yoganathan AP,Jo H

PA5-27322 was used in Western Blotting to test the role of novel flow-sensitive UBE2C (ubiquitin E2 ligase C) and microRNA-483-3p (miR-483) in flow-dependent aortic valve (AV) endothelial function and AV calcification.

Fri Mar 01 00:00:00 UTC 2019

miR-147b-mediated TCA cycle dysfunction and pseudohypoxia initiate drug tolerance to EGFR inhibitors in lung adenocarcinoma.

Nature metabolism

Zhang WC,Wells JM,Chow KH,Huang H,Yuan M,Saxena T,Melnick MA,Politi K,Asara JM,Costa DB,Bult CJ,Slack FJ

Published figure using VHL polyclonal antibody (Product # PA5-27322) in Western Blot

Mon Apr 01 00:00:00 UTC 2019

Bioinformatics

Protein Aliases: elongin binding protein; Protein G7; pVHL; VHL19; VHLp18(MEA); Von Hippel Lindau; von hippel-lindau; von Hippel-Lindau disease tumor suppressor; von Hippel-Lindau syndrome homolog; von Hippel-Lindau syndrome protein homolog; von Hippel-Lindau tumor suppressor, E3 ubiquitin protein ligase

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Gene Aliases: HRCA1; pVHL; RCA1; VHL; VHL1; Vhlh

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UniProt ID: (Human) P40337, (Mouse) P40338

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Entrez Gene ID: (Human) 7428, (Mouse) 22346

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ubiquitin-protein transferase activity protein binding transcription factor binding enzyme binding ubiquitin protein ligase activity protein complex binding

negative regulation of transcription from RNA polymerase II promoter cell morphogenesis regulation of transcription, DNA-templated proteolysis negative regulation of cell proliferation protein ubiquitination negative regulation of apoptotic process positive regulation of cell differentiation positive regulation of transcription, DNA-templated protein stabilization regulation of transcription from RNA polymerase II promoter in response to hypoxia negative regulation of transcription from RNA polymerase II promoter in response to hypoxia angiogenesis response to hypoxia type B pancreatic cell differentiation pancreatic A cell differentiation NLS-bearing protein import into nucleus proteasomal protein catabolic process protein transport protein catabolic process neuron differentiation extracellular matrix organization regulation of catecholamine metabolic process blood vessel endothelial cell migration negative regulation of endothelial cell differentiation protein heterooligomerization regulation of thymocyte apoptotic process negative regulation of thymocyte apoptotic process negative regulation of hypoxia-inducible factor-1alpha signaling pathway regulation of apoptotic signaling pathway

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