製品の概要

  • 製品名Anti-VCP antibody [5]
    VCP 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [5] to VCP
  • アプリケーション適用あり: IHC-P, IHC-Fr, Flow Cyt, ELISA, ICC, IP, ICC/IF, WBmore details
  • 種交差性
    交差種: Mouse, Rat, Sheep, Cow, Human
    交差が予測される動物種: Pig
  • 免疫原

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

    GGSVYTEDNDDDLYG


    (Peptide available as ab39788)

  • ポジティブ・コントロール
    • Whole cell lysate from cultured human B cells.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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 97 kDa.Can be blocked with Mouse VCP peptide (ab39788).

ターゲット情報

  • 機能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.
  • 関連疾患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.
  • 配列類似性Belongs to the AAA ATPase family.
  • 翻訳後修飾Phosphorylated by tyrosine kinases in response to T-cell antigen receptor activation (By similarity). Phosphorylated upon DNA damage, probably by ATM or ATR.
    ISGylated.
  • 細胞内局在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
  • 参照データベース
  • 別名
    • 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

Anti-VCP antibody [5] 画像

  • 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?)

    Image courtesy of an anonymous Abreview.

    See Abreview

  • 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 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.

  • 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 of VCP from CA46 cell lysate using 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 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.

  • 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.

  • 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.
  • 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.    

Anti-VCP antibody [5] (ab11433) 使用論文

This product has been referenced in:
  • Franz A  et al. Chromatin-associated degradation is defined by UBXN-3/FAF1 to safeguard DNA replication fork progression. Nat Commun 7:10612 (2016). Read more (PubMed: 26842564) »
  • Xue L  et al. Valosin-containing protein (VCP)-Adaptor Interactions are Exceptionally Dynamic and Subject to Differential Modulation by a VCP Inhibitor. Mol Cell Proteomics 15:2970-86 (2016). Read more (PubMed: 27406709) »

See all 24 Publications for this product

Product Wall

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
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Abcam user community

Verified customer

投稿 Oct 05 2016

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (u2os)
Specification u2os
Permeabilization Yes - 0.5% triton
Fixative Paraformaldehyde
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Abcam user community

Verified customer

投稿 Jan 08 2014

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (8%)
Sample Human Cell lysate - whole cell (u2os)
Specification u2os
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投稿 Jan 08 2014

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
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投稿 Dec 13 2013

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Sample Human Cell (U2OS)
Specification U2OS
Permeabilization Yes - 0.5% triton
Fixative Paraformaldehyde
Username

Dr. christophe lachaud

Verified customer

投稿 Jul 30 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Immuno-precipitation step Other - Dynabeads
Sample Human Cell lysate - whole cell (293 cells)
Specification 293 cells
Total protein in input 1000 µg
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投稿 Jul 24 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 10 µg
Gel Running Conditions Reduced Denaturing (8-16 %)
Sample Sheep Serum (Seminal plasma)
Specification Seminal plasma
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
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Mr. Guillaume TSIKIS

Verified customer

投稿 Jul 02 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry
Sample Mouse Cell (RAW264.7)
Specification RAW264.7
Fixative Paraformaldehyde
Permeabilization Yes - 0.1% Triton + 2% BSA in PBS
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 24°C
Username

Dr. Mahesh Shivananjappa

Verified customer

投稿 Apr 26 2013

Thank you for your response.
This is to let you know that I have placed a new order for you - for one vial of ab11433 as a free of charge replacement and the new order number is 1139141.
I hope the second vial will be intact without any damag...

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Thank you for contacting us.

Of the anti-VCP products which I had direct access to immunogen sequences, immunogen BLASTs show no homology with yeast. However there are four further products (ab76954, ab84385, ab138298, ab79037) which I do ...

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