p97/VCP Antibody

Western Blot: p97/VCP Antibody [NB100-1558] - Zymographic assay of gelatinase activity. Medium supernatants of 20ml culture medium (medium control) as well as 20 ml conditioned medium from 107 U937 cells, pMTH1-U937 cells and asCD11b-U937 cells in the absence or presence of 5nM TPA for 72 h, respectively, were 18-fold concentrated and subjected to SDS-PAGE containing 2 mg/ml of gelatine. Following incubation with MMP enzyme buffer, the gels were stained with 0.4% Coomassie blue and afterwards destained again to visualize the appearance of gelatinase activity by light bands against the dark background. The molecular weight markers on both sides of the gels indicate the size of the MMPs exhibiting gelatinase activities. Image collected and cropped by CiteAb from the following publication (//biosignaling.biomedcentral.com/articles/10.1186/1478-811X-10-13) licensed under a CC-BY license.

Western Blot: p97/VCP Antibody [NB100-1558] - Whole cell lysate (15 ug) from HeLa, 293T, Jurkat, mouse TCMK-1, and mouse NIH3T3 cells prepared using NETN lysis buffer. Antibody: Affinity purified rabbit antiVCP antibody used for WB at 0.1 ug/ml. Detection: Chemiluminescence with an exposure time of 1 second.

Western Blot: p97/VCP Antibody [NB100-1558] - Detection of human VCP by western blot of immunoprecipitates. Samples: Whole cell lysate (1.0 mg per IP reaction; 20% of IP loaded) from HeLa cells prepared using NETN lysis buffer. Antibodies: Affinity purified rabbit anti-VCP antibody NB100-1558 used for IP at 3 ug per reaction. VCP was also immunoprecipitated by rabbit anti-VCP antibody NB100-1557. For blotting immunoprecipitated VCP, NB100-1558 was used at 0.4 ug/ml. Detection: Chemiluminescence with an exposure time of 1 second.

Western Blot: p97/VCP Antibody [NB100-1558] - Zymographic assay of gelatinase activity. Medium supernatants of 20 ml culture medium (medium control) as well as 20 ml conditioned medium from 107 U937 cells, pMTH1-U937 cells & asCD11b-U937 cells in the absence or presence of 5nM TPA for 72 h, respectively, were 18-fold concentrated & subjected to SDS-PAGE containing 2 mg/ml of gelatine. Following incubation with MMP enzyme buffer, the gels were stained with 0.4% Coomassie blue & afterwards destained again to visualize the appearance of gelatinase activity by light bands against the dark background. The molecular weight markers on both sides of the gels indicate the size of the MMPs exhibiting gelatinase activities. Image collected & cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/22607136), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

• Reactivity: Hu, Mu, Rt, Ze, Po, Bv
• Applications: WB, ICC/IF, IHC, IP
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 0.2 mg/ml

p97/VCP Antibody Summary

• Immunogen: The immunogen recognized by this antibody maps to a region between residue 750 and the C-terminus (residue 806) of human Valosin-Containing Protein using the numbering given in entry NP_009057.1 (GeneID 7415).
• Predicted Species: Porcine (100%), Bovine (100%). Backed by our 100% Guarantee.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: VCP
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence 1:100 - 1:500
  • Immunohistochemistry
  • Immunohistochemistry-Frozen
  • Immunoprecipitation 2-5 ug/mg of lysate
  • Western Blot 1:2000 - 1:10000
• Application Notes: ICC/IF reactivity reported in (PMID: 23297223), IHC-Frozen reactivity reported in (PMID: 20833645).
• Theoretical MW: 89 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Control:

  p97/VCP Overexpression Lysate

Reactivity Notes

Zebrafish reactivity reported in scientific literature (PMID: 30010465). Mouse, Rat reactivity reported in scientific literature (PMID: 20833645).

Packaging, Storage & Formulations

• Storage: Store at 4C. Do not freeze.
• Buffer: TBS and 0.1% BSA
• Preservative: 0.09% Sodium Azide
• Concentration: 0.2 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for p97/VCP Antibody

• 15S Mg(2+)-ATPase p97 subunit
• IBMPFD
• IBMPFDMGC8560
• MGC131997
• p97
• TER ATPase
• TERA
• transitional endoplasmic reticulum ATPase
• valosin containing protein
• Valosin-containing proteinMGC148092
• VCP
• yeast Cdc48p homolog

Background

Valosin-containing protein (VCP or p97) belongs to the AAA (ATPase associated activities) family and acts as a molecular chaperone to a wide variety of cellular activities. Some of these activities include the alteration of both nuclear and mitotic golgi membranes, ubiquitin-proteasome dependent protein degradation, regulation of the NF-kappa b pathway, and extraction of membrane proteins. VCP has been shown to contain a substrate binding domain (N) and two conserved ATPase domains (D1 and D2). The three dimensional structure of VCP resembles that of a cylinder with the D1 and D2 stacked upon one another in a homo-hexameric ring formation. VCP is involved in the formation of neuronal inclusion bodies in neurodegenerative disease such as Parkinson's Disease.

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.

Publications for p97/VCP Antibody (NB100-1558)(12)

Publications using NB100-1558

• Saha I, Yuste-Checa P, Da Silva Padilha M et al. The AAA+ chaperone VCP disaggregates Tau fibrils and generates aggregate seeds in a cellular system Nature communications 2023-02-02 [PMID: 36732333]
• Garg J, Sporkova A, Hecker M, Korff T Tracing G-Protein-Mediated Contraction and Relaxation in Vascular Smooth Muscle Cell Spheroids Cells 2022-12-28 [PMID: 36611924]
• Leto DE, Morgens DW, Zhang L et al. Genome-wide CRISPR Analysis Identifies Substrate-Specific Conjugation Modules in ER-Associated Degradation Mol. Cell 2018-12-06 [PMID: 30581143] (Human)
• Kustermann M, Manta L, Paone C et al. Loss of the novel Vcp (valosin containing protein) interactor Washc4 interferes with autophagy-mediated proteostasis in striated muscle and leads to myopathy in vivo. Autophagy 2018-08-16 [PMID: 30010465] (WB, Zebrafish)
• Bersuker K, Peterson CWH, To M et al. A Proximity Labeling Strategy Provides Insights into the Composition and Dynamics of Lipid Droplet Proteomes. Dev. Cell 2017-12-19 [PMID: 29275994] (Human)
• Regensburger M, Turk M, Pagenstecher A et al. VCP-related multisystem proteinopathy presenting as early-onset Parkinson disease Neurology 2017-07-19 [PMID: 28724584] (Human)
• Olzmann JA, Richter CM, Kopito RR et al. Spatial regulation of UBXD8 and p97/VCP controls ATGL-mediated lipid droplet turnover Proc Natl Acad Sci U S A 2013-01-07 [PMID: 23297223] (ICC/IF, Human)
• Mandel K, Otte A, Hass R. Involvement of CD11b integrin in the alteration of metabolic factors after phorbol ester stimulation of human myeloid leukemia cells Cell Commun Signal 2012-07-11 [PMID: 22607136] (WB, Human)
• Christianson JC, Olzmann JA, Shaler TA et al. Defining human ERAD networks through an integrative mapping strategy. Nature Cell Biology. 2011-11-27 [PMID: 22119785]
• Clemen CS, Tangavelou K, Strucksberg KH et al. Strumpellin is a novel valosin-containing protein binding partner linking hereditary spastic paraplegia to protein aggregation diseases. Brain 2010-10-01 [PMID: 20833645] (IHC-Fr, Rat, Mouse)
• Leto D, Morgens D, Zhang L et al. Parallel genome-wide CRISPR analysis identifies a role for heterotypic ubiquitin chains in ER-associated degradation bioRxiv 2018-06-17 (WB, Human)
• Berschneider K Connecting the functions of the proteasome and mitochondria in the lung Thesis 2016-01-01 (WB, Mouse)

FAQs for p97/VCP Antibody (NB100-1558). (Showing 1 - 1 of 1 FAQs).

1. I used the p97/VCP antibody (catalog # NB100-1558) and would now like to use for ICC. Is there a recommended dilution? In the Data Sheet can be found on the ICC not specified.
   • For the product NB100-1558, p97/VCP Antibody, the recommended starting dilution for ICC/IF is 1:100 to 1:500. Please note that we always advise customers to investigate the optimal dilution for them, as this will depend on the particular samples/reagents/protocols being used.

Bioinformatics

• Gene Symbol: VCP
• Entrez:
  • 7415(Human)
  • 67459(Mouse)
• Uniprot:
  • P55072(Human)