Anti-VCP antibody [5] (ab11433)

Reviews (24)Q&A (6)References (77)

Overview

  • Product name

    Anti-VCP antibody [5]
    See all VCP primary antibodies

  • Description

    Mouse monoclonal [5] to VCP

  • Host species

    Mouse

  • Tested applications

    Suitable for: IHC-P, IHC-Fr, Flow Cyt, ELISA, ICC, IP, ICC/IF, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Sheep, Cow, Human
    Predicted to work with: Pig

  • Immunogen

    Synthetic peptide corresponding to Human VCP aa 792-806.
    Sequence:

    GGSVYTEDNDDDLYG


    (Peptide available as ab39788)

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • Positive control

    • Whole cell lysate from cultured human B cells.

  • General notes

    Abcam is committed to meeting high quality standards of ethical manufacturing and has decided to discontinue this product by June 2020 as it has been generated by the ascites method. We are sorry for any inconvenience this may cause. We suggest ab109240 or ab110308 as possible replacements.

     

Properties

Applications

Our Abpromise guarantee covers the use of ab11433 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
IHC-P 1/500.
IHC-Fr 1/500.
Flow Cyt 1/400.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
ELISA Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
ICC/IF 1/20 - 1/200.
WB 1/2000. Detects a band of approximately 89 kDa.Can be blocked with Mouse VCP peptide (ab39788).

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

    see all

  • 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

  • Western blot - Anti-VCP antibody [5] (ab11433)
    Western blot - Anti-VCP antibody [5] (ab11433)Image courtesy of an anonymous Abreview.
    Anti-VCP antibody [5] (ab11433) at 1/2000 dilution + whole cell lysate prepared from mouse embryonic fibroblasts at 20 µg

    Secondary
    Alexa-Fluor 680 conjugated goat anti-mouse polyclonal at 1/10000 dilution

    Observed band size: 97 kDa
    why is the actual band size different from the predicted?

    See Abreview

  • Immunocytochemistry/ Immunofluorescence - Anti-VCP antibody [5] (ab11433)
    Immunocytochemistry/ Immunofluorescence - Anti-VCP antibody [5] (ab11433)
    Immunofluorescent analysis of VCP using VCP Monoclonal antibody (5) ab11433 shows staining in HeLa cells. VCP staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing VCP ab11433 at a dilution of 1:20-1:200 over night at 4 ?C washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VCP antibody [5] (ab11433)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VCP antibody [5] (ab11433)
    Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human colon carcinoma tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:1000 with a mouse monoclonal antibody recognizing Anti-VCP ab11433 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Flow Cytometry - Anti-VCP antibody [5] (ab11433)
    Flow Cytometry - Anti-VCP antibody [5] (ab11433)This image is courtesy of an Abreview by Mahesh Shivananjappa.
    ab24762 detecting VCP in Human platelets by Flow Cytometry. Platelets were isolated by spinning Platelet rich plasma on Histopaque. Cells were fixed in PFA and permeabilized in 0.1% Triton-X100 in 2% BSA for 30 minutes. The primary antibody was diluted 1/400 in 2% BSA in PBS and incubated with the sample for 18 hours at 4°C. An Alexa Fluor® 488-conjugated Goat anti-Mouse IgG (H+L) antibody, diluted 1/500 was used as the secondary antibody.

    Abbreviations in the figure:
    P : Permeabilized;
    US Unstained, Red Peak;
    IgG Rb: IgG Mouse (Isotype Control), Blue Peak;
    VCP: Green Peak.

    See Abreview

  • Western blot - Anti-VCP antibody [5] (ab11433)
    Western blot - Anti-VCP antibody [5] (ab11433)

    Western blot of VCP from CA46 cell lysate using ab11433.

  • Immunocytochemistry/ Immunofluorescence - Anti-VCP antibody [5] (ab11433)
    Immunocytochemistry/ Immunofluorescence - Anti-VCP antibody [5] (ab11433)
    Immunofluorescent analysis of VCP using VCP Monoclonal antibody (5) ab11433 shows staining in C6 glioma cells. VCP staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing VCP ab11433 at a dilution of 1:20-1:200 over night at 4 ?C washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VCP antibody [5] (ab11433)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VCP antibody [5] (ab11433)
    Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human brain tissue tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:1000 with a mouse monoclonal antibody recognizing Anti-VCP ab11433 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Immunocytochemistry/ Immunofluorescence - Anti-VCP antibody [5] (ab11433)
    Immunocytochemistry/ Immunofluorescence - Anti-VCP antibody [5] (ab11433)
    Immunofluorescent analysis of VCP using VCP Monoclonal antibody (5) ab11433 shows staining in WiDr colon carcinoma cells. VCP staining (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing VCP ab11433 at a dilution of 1:20-1:200 over night at 4 ?C washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VCP antibody [5] (ab11433)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VCP antibody [5] (ab11433)
    ab11433 (1µg/ml) staining VCP in human left ventricle using an automated system (DAKO Autostainer Plus). Using this protocol there is strong nuclear, cytoplasmic and cell membrane staining of cardiomyocutes and smooth muscle cells of cardiac artery.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
  • Immunocytochemistry/ Immunofluorescence - Anti-VCP antibody [5] (ab11433)
    Immunocytochemistry/ Immunofluorescence - Anti-VCP antibody [5] (ab11433)This image is courtesy of an anonymous Abreview.

    ab11433 staining VCP in Human H1299 cells by Immunocytochemistry/ Immunofluorescence. The cells were paraformaldehyde fixed, permeabilized in 0.1% Triton X-100  and were blocked in 10% serum at 25°C for 1 hour. Primary antibody was used at 1/1000 dilution (PBS) and incubated with sample for 1 hour at 37°C. An Alexa Fluor® 488 conjugated Goat polyclonal to mouse IgG was used as secondary at 1/1000 dilution. Image shows green color staining with ab11433 and nuclei were stained blue with DAPI.    

References

This product has been referenced in:

  • Wolff G  et al. Diet-dependent function of the extracellular matrix proteoglycan Lumican in obesity and glucose homeostasis. Mol Metab 19:97-106 (2019). Read more (PubMed: 30409703) »
  • Glantschnig C  et al. A miR-29a-driven negative feedback loop regulates peripheral glucocorticoid receptor signaling. FASEB J 33:5924-5941 (2019). Read more (PubMed: 30742779) »
See all 77 Publications for this product

Publishing research using ab11433? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-10 of 30 Abreviews or Q&A

Western blot abreview for Anti-VCP antibody [5]

 Good
Abreviews
abreview image
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
Read More

Nan Xin

Verified customer

Submitted May 15 2019

Western blot abreview for Anti-VCP antibody [5]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Vascular smooth muscle cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
Vascular smooth muscle cells
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Read More

Abcam user community

Verified customer

Submitted Jun 25 2018

Immunocytochemistry/ Immunofluorescence abreview for Anti-VCP antibody [5]

 Good
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HAP1)
Permeabilization
Yes - 0.5% Triton 5 minutes
Specification
HAP1
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Feb 15 2018

Western blot abreview for Anti-VCP antibody [5]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (MEF cells)
Gel Running Conditions
Reduced Denaturing (4-12% Gradient Gel)
Loading amount
20 µg
Specification
MEF cells
Blocking step
Licor Blocking Buffer as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted May 03 2017

Western blot abreview for Anti-VCP antibody [5]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HUVEC)
Gel Running Conditions
Reduced Denaturing
Loading amount
10 µg
Specification
HUVEC
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Dr. Juan Rodríguez Vita

Verified customer

Submitted Mar 31 2017

Western blot abreview for Anti-VCP antibody [5]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (3T3 Fibroblasts)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
10 µg
Treatment
TGF-beta for 20 min or 24 h
Specification
3T3 Fibroblasts
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Mar 07 2017

Western blot abreview for Anti-VCP antibody [5]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (U2OS cells)
Gel Running Conditions
Reduced Denaturing (4-12% Gradient Gel)
Loading amount
50 µg
Specification
U2OS cells
Blocking step
Licor Blocking Buffer as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Oct 05 2016

Immunocytochemistry/ Immunofluorescence abreview for Anti-VCP [5] antibody

 Good
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (u2os)
Specification
u2os
Permeabilization
Yes - 0.5% triton
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Jan 08 2014

Western blot abreview for Anti-VCP [5] antibody

 Good
Abreviews
abreview image
Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (8%)
Sample
Human Cell lysate - whole cell (u2os)
Specification
u2os
Read More

Abcam user community

Verified customer

Submitted Jan 08 2014

Immunocytochemistry/ Immunofluorescence abreview for Anti-VCP [5] antibody

 Good
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: RT°C
Sample
Human Cell (Human primary fibroblasts)
Specification
Human primary fibroblasts
Permeabilization
Yes - 0.2% Triton-X100
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

Submitted Dec 13 2013

1-10 of 30 Abreviews or Q&A

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