Name: Anti-VCP antibody [EPR3307(2)]
SKU: ab109240
Rating: 4 (1 reviews)

Rabbit Recombinant Monoclonal VCP antibody. Suitable for IHC-P, IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples. Cited in 28 publications.

Images

Western blot - Anti-VCP antibody [EPR3307(2)] (AB109240), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IHC-P	IP	WB	ICC/IF	Flow Cyt (Intra)
Human	Tested	Tested	Tested	Tested	Tested
Mouse	Expected	Expected	Tested	Expected	Expected
Rat	Expected	Expected	Tested	Expected	Expected

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100 - 1/250	Notes Perform heat-mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/10 - 1/100	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/10000	Notes For unpurifed use at 1/10000-1/50000.
Species Rat	Dilution info 1/10000	Notes For unpurifed use at 1/10000-1/50000.
Species Human	Dilution info 1/10000 - 1/50000	Notes For unpurifed use at 1/10000-1/50000.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/500	Notes For unpurified use at 1/100 - 1/250.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/300	Notes For unpurified use at 1/10 - 1/100. Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Associated Products

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Target data

See full target information VCP

Function

Necessary for the fragmentation of Golgi stacks during mitosis and for their reassembly after mitosis. Involved in the formation of the transitional endoplasmic reticulum (tER). The transfer of membranes from the endoplasmic reticulum to the Golgi apparatus occurs via 50-70 nm transition vesicles which derive from part-rough, part-smooth transitional elements of the endoplasmic reticulum (tER). Vesicle budding from the tER is an ATP-dependent process. The ternary complex containing UFD1, VCP and NPLOC4 binds ubiquitinated proteins and is necessary for the export of misfolded proteins from the ER to the cytoplasm, where they are degraded by the proteasome. The NPLOC4-UFD1-VCP complex regulates spindle disassembly at the end of mitosis and is necessary for the formation of a closed nuclear envelope. Regulates E3 ubiquitin-protein ligase activity of RNF19A. Component of the VCP/p97-AMFR/gp78 complex that participates in the final step of the sterol-mediated ubiquitination and endoplasmic reticulum-associated degradation (ERAD) of HMGCR. Mediates the endoplasmic reticulum-associated degradation of CHRNA3 in cortical neurons as part of the STUB1-VCP-UBXN2A complex (PubMed:26265139). Involved in endoplasmic reticulum stress-induced pre-emptive quality control, a mechanism that selectively attenuates the translocation of newly synthesized proteins into the endoplasmic reticulum and reroutes them to the cytosol for proteasomal degradation (PubMed:26565908). Involved in clearance process by mediating G3BP1 extraction from stress granules (PubMed:29804830, PubMed:34739333). Also involved in DNA damage response: recruited to double-strand breaks (DSBs) sites in a RNF8- and RNF168-dependent manner and promotes the recruitment of TP53BP1 at DNA damage sites (PubMed:22020440, PubMed:22120668). Recruited to stalled replication forks by SPRTN: may act by mediating extraction of DNA polymerase eta (POLH) to prevent excessive translesion DNA synthesis and limit the incidence of mutations induced by DNA damage (PubMed:23042605, PubMed:23042607). Together with SPRTN metalloprotease, involved in the repair of covalent DNA-protein cross-links (DPCs) during DNA synthesis (PubMed:32152270). Involved in interstrand cross-link repair in response to replication stress by mediating unloading of the ubiquitinated CMG helicase complex (By similarity). Mediates extraction of PARP1 trapped to chromatin: recognizes and binds ubiquitinated PARP1 and promotes its removal (PubMed:35013556). Required for cytoplasmic retrotranslocation of stressed/damaged mitochondrial outer-membrane proteins and their subsequent proteasomal degradation (PubMed:16186510, PubMed:21118995). Essential for the maturation of ubiquitin-containing autophagosomes and the clearance of ubiquitinated protein by autophagy (PubMed:20104022, PubMed:27753622). Acts as a negative regulator of type I interferon production by interacting with RIGI: interaction takes place when RIGI is ubiquitinated via 'Lys-63'-linked ubiquitin on its CARD domains, leading to recruit RNF125 and promote ubiquitination and degradation of RIGI (PubMed:26471729). May play a role in the ubiquitin-dependent sorting of membrane proteins to lysosomes where they undergo degradation (PubMed:21822278). May more particularly play a role in caveolins sorting in cells (PubMed:21822278, PubMed:23335559). By controlling the steady-state expression of the IGF1R receptor, indirectly regulates the insulin-like growth factor receptor signaling pathway (PubMed:26692333).

Alternative names

HEL-220, HEL-S-70, VCP, Transitional endoplasmic reticulum ATPase, TER ATPase, 15S Mg(2+)-ATPase p97 subunit, Valosin-containing protein

Recommended products

Rabbit Recombinant Monoclonal VCP antibody. Suitable for IHC-P, IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples. Cited in 28 publications.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR3307(2)
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle, Stable for 12 months at -20°C

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

The VCP protein also known as p97 or valosin-containing protein is a significant ATPase. It is involved mechanically in a range of cellular activities. VCP has a mass of approximately 97 kDa and is ubiquitously expressed in many tissues. It plays a role in protein homeostasis particularly in the degradation process of misfolded proteins via the ubiquitin-proteasome system. This protein comprises the D1 and D2 ATPase domains that are important for its unfolding activities.

Biological function summary

The VCP protein operates as a vital part of cellular machinery and functions in various complexes. It assists in protein quality control by participating in processes such as ER-associated degradation (ERAD) where it retrotranslocates misfolded proteins from the endoplasmic reticulum for degradation. VCP also contributes to the regulation of endocytosis autophagy and cell cycle control. These functions are important in maintaining cellular integrity.

Pathways

VCP integrates into cellular processes by engaging in essential pathways like the ubiquitin-proteasome system and autophagy. It associates with proteins such as UFD1 and NPL4 in these pathways illustrating its multifaceted role in cellular regulation. VCP's action is closely tied with the p97 protein complexes where it influences protein degradation and recycling thereby controlling protein turnover.

Associated diseases and disorders

There are findings linking VCP to conditions such as inclusion body myopathy and Paget's disease of bone. Its malfunction due to mutations can trigger these diseases affecting muscle and bone tissues respectively. VCP's connection to other proteins like Parkin and Optineurin in these disorders highlights the protein's relevance in neurodegenerative and skeletal disease pathways emphasizing its pivotal role in maintaining cellular health.

Product promise

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10 product images

Western blot - Anti-VCP antibody [EPR3307(2)] (ab109240)
Blocking and diluting buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-VCP antibody [EPR3307(2)] (ab109240) at 1/10000 dilution
Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 20 µg
Lane 2: A549 (Human lung carcinoma epithelial cell) whole cell lysates at 20 µg
Lane 3: C6 (Rat glial tumor glial cell) whole cell lysates) at 20 µg
Lane 4: NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates at 20 µg
Lane 5: RAW264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates at 20 µg
Lane 6: SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysates at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 112 kDa, 130 kDa, 151 kDa, 217 kDa, 43 kDa, 469 kDa, 47 kDa, 60 kDa, 65 kDa, 89 kDa, 92 kDa
Observed band size: 89 kDa
Immunoprecipitation - Anti-VCP antibody [EPR3307(2)] (ab109240)
ab109240 (purified) at 1:20 dilution (2ug) immunoprecipitating VCP in HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate.

Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug

Lane 2 (+): ab109240 & HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate

Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab109240 in HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate

For western blotting, VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used for detection at 1:1000 dilution.

Blocking and diluting buffer: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-VCP antibody [EPR3307(2)] (ab109240)
Predicted band size: 89 kDa
Observed band size: 89 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VCP antibody [EPR3307(2)] (ab109240)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast carcinoma tissue sections labeling VCP with purified ab109240 at 1:250 dilution (1.4 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, pH9.0. Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.
Immunocytochemistry/ Immunofluorescence - Anti-VCP antibody [EPR3307(2)] (ab109240)
Immunofluorescent staining of HeLa cells using unpurified ab109240 at 1/100 dilution.
Flow Cytometry (Intracellular) - Anti-VCP antibody [EPR3307(2)] (ab109240)
Intracellular Flow Cytometry analysis of HL-60 (Human acute promyelocytic leukemia promyeloblast) cells labeling VCP with purified ab109240 at 1/300 dilution (1 ug/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluorr^® 488) secondary antibody was used at 1/2000 dilution. Isotype control - 90% methanol. Unlabeled control - Rabbit monoclonal IgG (Black).Cell without incubation with primary antibody and secondary antibody (Blue).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VCP antibody [EPR3307(2)] (ab109240)
Immunocytochemistry/ Immunofluorescence analysis of MCF-7 (Human breast adenocarcinoma epithelial cell) cells labeling VCP with Purified ab109240 at 1:500 dilution (0.7μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1:200 (2.5 μg/ml). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat anti rabbit IgG(Alexa Fluor^® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VCP antibody [EPR3307(2)] (ab109240)
Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using unpurified ab109240 at 1/100 dilution.
Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
Western blot - Anti-VCP antibody [EPR3307(2)] (ab109240)
All lanes: Western blot - Anti-VCP antibody [EPR3307(2)] (ab109240) at 1/10000 dilution
Lane 1: MCF7 cell lysate at 10 µg/mL
Lane 2: HeLa cell lysate at 10 µg/mL
Lane 3: A549 cell lysate at 10 µg/mL
Lane 4: SH-SY5Y cell lysate at 10 µg/mL
Predicted band size: 89 kDa
Flow Cytometry (Intracellular) - Anti-VCP antibody [EPR3307(2)] (ab109240)
Overlay histogram showing HL60 cells stained with unpurified ab109240 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab109240, 1/100) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HL60 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Western blot - Anti-VCP antibody [EPR3307(2)] (ab109240)
ab109240 was shown to react with VCP in wild-type U2OS cells in Western blot with loss of signal observed in a VCP siRNA knockdown cell line. Cell lysates from wild-type U2OS transfected with either scrambled siRNA or VCP siRNA were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab109240 overnight at 4 °C at a 1/50000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
All lanes: Western blot - Anti-VCP antibody [EPR3307(2)] (ab109240) at 1/50000 dilution
Lane 1: Wild-type U2OS transfected with scrambled siRNA control lysate at 10 µg
Lane 2: U2OS transfected with siRNA specifically targeting VCP cell lysate at 10 µg