Overview

  • Product name

    Anti-VCP antibody [EPR3307(2)]
    See all VCP primary antibodies
  • Description

    Rabbit monoclonal [EPR3307(2)] to VCP
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Human VCP aa 1-100.

  • Positive control

    • MCF-7 cell lysate, HeLa whole cell lysate (ab150035), A549 cell lysate; SH-SY5Y cell lysate; human breast carcinoma tissue; HeLa cells
  • General 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab109240 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/10000. Predicted molecular weight: 89 kDa.

For unpurifed use at 1/10000-1/50000.

IP 1/10 - 1/100.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Flow Cyt 1/300.

For unpurified use at 1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/500.

For unpurified use at 1/100 - 1/250.

Target

  • 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 UFD1L, 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-UFD1L-VCP complex regulates spindle disassembly at the end of mitosis and is necessary for the formation of a closed nuclear envelope (By similarity). Regulates E3 ubiquitin-protein ligase activity of RNF19A.
  • Involvement in disease

    Defects in VCP are the cause of inclusion body myopathy with early-onset Paget disease and frontotemporal dementia (IBMPFD) [MIM:167320]; also known as muscular dystrophy, limb-girdle, with Paget disease of bone or pagetoid amyotrophic lateral sclerosis or pagetoid neuroskeletal syndrome or lower motor neuron degeneration with Paget-like bone disease. IBMPFD features adult-onset proximal and distal muscle weakness (clinically resembling limb girdle muscular dystrophy), early-onset Paget disease of bone in most cases and premature frontotemporal dementia.
  • Sequence similarities

    Belongs to the AAA ATPase family.
  • Post-translational
    modifications

    Phosphorylated by tyrosine kinases in response to T-cell antigen receptor activation (By similarity). Phosphorylated upon DNA damage, probably by ATM or ATR.
    ISGylated.
  • Cellular localization

    Cytoplasm > cytosol. Nucleus. Present in the neuronal hyaline inclusion bodies specifically found in motor neurons from amyotrophic lateral sclerosis patients. Present in the Lewy bodies specifically found in neurons from Parkinson disease patients.
  • Information by UniProt
  • Database links

  • Alternative names

    • 15S Mg(2+) ATPase p97 subunit antibody
    • 15S Mg(2+)-ATPase p97 subunit antibody
    • ALS14 antibody
    • ATPase p97 antibody
    • CDC48 antibody
    • IBMPFD antibody
    • MGC131997 antibody
    • MGC148092 antibody
    • MGC8560 antibody
    • p97 antibody
    • TER ATPase antibody
    • TERA antibody
    • TERA_HUMAN antibody
    • Transitional endoplasmic reticulum ATPase antibody
    • Valosin containing protein antibody
    • Valosin-containing protein antibody
    • VCP antibody
    • Yeast Cdc48p homolog antibody
    see all

Images

  • All lanes : Anti-VCP antibody [EPR3307(2)] (ab109240) at 1/10000 dilution (purified)

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
    Lane 2 : A549 (Human lung carcinoma epithelial cell) whole cell lysates
    Lane 3 : C6 (Rat glial tumor glial cell) whole cell lysates)
    Lane 4 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates
    Lane 5 : RAW264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates
    Lane 6 : SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 89 kDa
    Observed band size: 89 kDa



    Blocking and diluting buffer: 5% NFDM/TBST

  • 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 Perform heat mediated antigen retrieval 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.

  • Immunofluorescent staining of HeLa cells using unpurified ab109240 at 1/100 dilution.

  • 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 (ab172730) instead of ab109240 in HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate

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

    Blocking and diluting buffer: 5% NFDM/TBST.

  • 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 Fluor® 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).

  • 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 Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). 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.

  • 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.

  • All lanes : Anti-VCP antibody [EPR3307(2)] (ab109240) at 1/10000 dilution (unpurified)

    Lane 1 : MCF7 cell lysate
    Lane 2 : HeLa cell lysate
    Lane 3 : A549 cell lysate
    Lane 4 : SH-SY5Y cell lysate

    Lysates/proteins at 10 µg/ml per lane.

    Predicted band size: 89 kDa

  • 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 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. 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.

References

This product has been referenced in:

  • Kaneko T  et al. Identification and characterization of a large family of superbinding bacterial SH2 domains. Nat Commun 9:4549 (2018). Read more (PubMed: 30382091) »
  • Wu Y  et al. Sp110 enhances macrophage resistance toMycobacterium tuberculosisvia inducing endoplasmic reticulum stress and inhibiting anti-apoptotic factors. Oncotarget 8:64050-64065 (2017). Read more (PubMed: 28969051) »
See all 12 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Western blot
Sample
Caenorhabditis elegans Tissue lysate - whole (whole worm lysate)
Gel Running Conditions
Reduced Denaturing (4-15%)
Loading amount
20 µg
Specification
whole worm lysate
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Nan Xin

Verified customer

Submitted May 08 2019

Answer

Thank you for contacting us.

I have gathered the information which you have requested:
ab109240: The immunogen for this product is located at the N-terminal of VCP. This product is guaranteed in WB and IP
ab79037: I have been informed by the lab that they do not know the specific epitope site for this antibody.
ab19444: This product is created from the full length peptide, epitope mapping has not been performed so we cannot guarantee that this does not recognize the C-terminal of VCP.


I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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