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Anti vcp

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Anti-VCP is a primary antibody that recognizes the VCP (Valosin-Containing Protein) protein. VCP is an essential protein involved in various cellular processes, including protein degradation, membrane fusion, and cell cycle regulation. The Anti-VCP antibody can be used to detect and study the expression and localization of VCP in biological samples.

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Lab products found in correlation

Anti ha, by Cell Signaling Technology (3 mentions) Anti ataxin 3, by Merck Group (3 mentions) Peroxidase conjugated secondary antibodies goat anti mouse and goat anti rabbit, by Jackson ImmunoResearch (2 mentions) Anti myc, by Santa Cruz Biotechnology (2 mentions) Anti tubulin, by Merck Group (1 mentions) Anti enos, by Santa Cruz Biotechnology (1 mentions) Anti inos, by Abcam (1 mentions) Anti β actin, by Merck Group (1 mentions) Anti cleaved parp, by Cell Signaling Technology (1 mentions) Pvdf membrane, by Merck Group (1 mentions) Complete protease inhibitor cocktail tablet, by Roche (1 mentions) Immobilon western kit, by Merck Group (1 mentions) Anti survivin, by Cell Signaling Technology (1 mentions) Chemidoc xrs system, by Bio-Rad (1 mentions) Hrp linked anti mouse igg, by Merck Group (1 mentions) Hrp linked anti rabbit igg, by Merck Group (1 mentions) Hrp conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) Bis tris gel, by Thermo Fisher Scientific (1 mentions) Anti gfp clone b 2, by Santa Cruz Biotechnology (1 mentions) Anti bag2, by Merck Group (1 mentions)

7 protocols using anti vcp

1

Western Blot Antibody Protocol

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Anti-Ataxin-3 (mouse monoclonal 1H9, 1:500–1000; Millipore) (rabbit polyclonal, 1:15000; (Paulson et al., 1997 (link))), anti-tubulin (mouse monoclonal T5168, 1:10000; Sigma-Aldrich), anti-VCP (rabbit monoclonal, 1:1000; Cell Signaling Technology), anti-HA (rabbit monoclonal, 1:500–1000; Cell Signaling Technology), anti-lamin (mouse monoclonal ADL84.12-5, 1:1000; Developmental Studies Hybridoma Bank), peroxidase conjugated secondary antibodies (goat antimouse and goat anti-rabbit, 1:5000; Jackson Immunoresearch).
Ristic G., Sutton J.R., Libohova K, & Todi S.V. (2018). Toxicity and aggregation of the polyglutamine disease protein, ataxin-3 is regulated by its binding to VCP/p97 in Drosophila melanogaster. Neurobiology of disease, 116, 78-92.
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2

Protein Expression Analysis in Mouse Cardiac Tissues

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mRNA was extracted from mouse heart tissues and qPCR was performed as described previously49 (link)54 (link). Protein extraction and subcellular fractions were performed as described previously49 (link)54 (link). Targeted proteins including VCP, eNOS and iNOS were detected by western blotting as previously described15 (link)18 (link). The primary antibodies used for western blotting were anti-VCP (catalog no. 2648S, rabbit, dilution of 1:1000, Cell Signaling Technology), anti-iNOS (catalog no. ab136918, rabbit, dilution of 1:1000, Abcam), and anti-eNOS (catalog no. Sc-654, rabbit, dilution of 1:1000, Santa Cruz Biotechnology). Anti-GAPDH (Catalog no. ab9484, mouse, dilution of 1:10,000, Abcam) and anti- voltage-dependent anion-selective channel protein 1(VDAC) (Catalog no. 4866, rabbit, dilution of 1:5,000, Cell Signaling Technology) were used as loading controls.
Lizano P., Rashed E., Stoll S., Zhou N., Wen H., Hays T.T., Qin G., Xie L.H., Depre C, & Qiu H. (2017). The valosin-containing protein is a novel mediator of mitochondrial respiration and cell survival in the heart in vivo. Scientific Reports, 7, 46324.
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3

Western Blotting Analysis of Apoptosis Markers

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Anti‐cleaved‐PARP, anti‐VCP and anti‐survivin were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti‐β‐actin, HRP‐linked anti‐mouse IgG and HRP‐linked anti‐rabbit IgG were purchased from Sigma‐Aldrich. Cells were immediately harvested and lysed with RIPA buffer (25 mM HEPES, 1.5% TX‐100, 1% sodium deoxycholate, 0.1% SDS, 500 mM NaCl, 5 mM EDTA, 50 mM NaF, 0.1 M Na3VO4 and cOmplete Protease Inhibitor Cocktail Tablets [Roche, Mannheim, Germany]; pH 7.8). The lysates were centrifuged at 13 000 g for 15 min to remove the insoluble fraction. Equal amounts of total protein were subsequently separated by SDS‐polyacrylamide gel electrophoresis before being transferred onto a PVDF membrane (Millipore, Billerica, MA, USA). The membrane was probed with the indicated antibodies. The chemiluminescence signal was detected using an Immobilon Western kit (Millipore) and ChemiDoc XRS+ System (Bio‐Rad, Hercules, CA, USA).
Shikata Y., Yoshimaru T., Komatsu M., Katoh H., Sato R., Kanagaki S., Okazaki Y., Toyokuni S., Tashiro E., Ishikawa S., Katagiri T, & Imoto M. (2017). Protein kinase A inhibition facilitates the antitumor activity of xanthohumol, a valosin‐containing protein inhibitor. Cancer Science, 108(4), 785-794.
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4

Protein Expression Analysis in Insect Larvae

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Third instar larvae were dissected and muscle preparations were immediately transferred into 5× SDS sample buffer and denatured by boiling for 10 min. Proteins were resolved by SDS-PAGE on a 4–12% Bis-Tris gel (Life Technologies), transferred to a nitrocellulose membrane, immunoblotted with primary and HRP-conjugated secondary antibodies (Life Technologies) and detected using an ECL chemi-luminescent reagent (Life Technologies). The following primary antibodies were used: anti-VCP (Cell Signaling, Danvers, MA) at 1:1000, anti- GRP78/HSPA5 (Thermo Scientific) at 1:2500, insect anti-Cathepsin L (R&D Systems, Minneapolis, MN) at 1:1000, and anti-Tubulin E7-c (Developmental Studies Hybridoma Bank, University of Iowa, IA) at 1:10,000. Secondary HRP conjugated antibodies (GE Lifesciences, Pittsburgh, PA) were used at 1:5000.
Johnson A.E., Shu H., Hauswirth A.G., Tong A, & Davis G.W. (2015). VCP-dependent muscle degeneration is linked to defects in a dynamic tubular lysosomal network in vivo. eLife, 4, e07366.
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5

Antibody Immunodetection in Ataxin-3 Studies

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Anti-Ataxin-3: mouse monoclonal 1H9, 1:500–1000; Millipore; rabbit polyclonal, 1:15000 (Paulson et al., 1997 (link)). Anti-HA: rabbit monoclonal, 1:500–1000; Cell Signaling Technology. Anti-Myc: mouse monoclonal 9E10, 1:500–1000; Santa Cruz Biotechnology. Anti-VCP: rabbit monoclonal, 1:500–1:1000; Cell Signaling Technology. Peroxidase-conjugated secondary antibodies: goat anti-mouse and goat anti-rabbit, 1:5000; Jackson ImmunoResearch.
Johnson S.L., Libohova K., Blount J.R., Sujkowski A.L., Prifti M.V., Tsou W.L, & Todi S.V. (2021). Targeting the VCP-binding motif of ataxin-3 improves phenotypes in Drosophila models of Spinocerebellar Ataxia Type 3. Neurobiology of disease, 160, 105516.
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6

Antibody-Based Protein Detection in Cells

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The following antibodies were used: anti-GFP (B-2 clone; Santa Cruz; sc-9996), anti-HSP70 (Enzo Life Sciences; N15F2-5), anti-BAG6 (Cell Signaling Technology, 8523), anti-VCP (Cell Signaling Technology; 2649), anti-BAG2 (Sigma Alrich; HPA018862), anti-PRPF19 (Abcam; ab27692), anti-POLR1A (Santa Cruz; sc-48385), anti-POLR3A (D5Y2D; Cell Signaling Technology, 12825).
Hildebrandt A., Alanis-Lobato G., Voigt A., Zarnack K., Andrade-Navarro M.A., Beli P, & König J. (2017). Interaction profiling of RNA-binding ubiquitin ligases reveals a link between posttranscriptional regulation and the ubiquitin system. Scientific Reports, 7, 16582.
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7

Antibody Immunodetection in Ataxin-3 Studies

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Anti-Ataxin-3: mouse monoclonal 1H9, 1:500–1000; Millipore; rabbit polyclonal, 1:15000 (Paulson et al., 1997 (link)). Anti-HA: rabbit monoclonal, 1:500–1000; Cell Signaling Technology. Anti-Myc: mouse monoclonal 9E10, 1:500–1000; Santa Cruz Biotechnology. Anti-VCP: rabbit monoclonal, 1:500–1:1000; Cell Signaling Technology. Peroxidase-conjugated secondary antibodies: goat anti-mouse and goat anti-rabbit, 1:5000; Jackson ImmunoResearch.
Johnson S.L., Libohova K., Blount J.R., Sujkowski A.L., Prifti M.V., Tsou W.L, & Todi S.V. (2021). Targeting the VCP-binding motif of ataxin-3 improves phenotypes in Drosophila models of Spinocerebellar Ataxia Type 3. Neurobiology of disease, 160, 105516.
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