Anti-Vimentin 抗体 [EPR3776] - Cytoskeleton Marker (ab92547)

製品の概要

製品の特性

関連製品

アプリケーション

Our Abpromise guarantee covers the use of ab92547 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
ICC/IF 1/250.
WB 1/1000 - 1/5000. Predicted molecular weight: 54 kDa.
Flow Cyt 1/50.

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

IHC-P 1/200 - 1/500.
IHC-Fr 1/100 - 1/500.

ターゲット情報

  • 機能Vimentins are class-III intermediate filaments found in various non-epithelial cells, especially mesenchymal cells. Vimentin is attached to the nucleus, endoplasmic reticulum, and mitochondria, either laterally or terminally.
    Involved with LARP6 in the stabilization of type I collagen mRNAs for CO1A1 and CO1A2.
  • 組織特異性Highly expressed in fibroblasts, some expression in T- and B-lymphocytes, and little or no expression in Burkitt's lymphoma cell lines. Expressed in many hormone-independent mammary carcinoma cell lines.
  • 関連疾患Cataract 30
  • 配列類似性Belongs to the intermediate filament family.
  • ドメインThe central alpha-helical coiled-coil rod region mediates elementary homodimerization.
    The [IL]-x-C-x-x-[DE] motif is a proposed target motif for cysteine S-nitrosylation mediated by the iNOS-S100A8/A9 transnitrosylase complex.
  • 翻訳後修飾Filament disassembly during mitosis is promoted by phosphorylation at Ser-55 as well as by nestin (By similarity). One of the most prominent phosphoproteins in various cells of mesenchymal origin. Phosphorylation is enhanced during cell division, at which time vimentin filaments are significantly reorganized. Phosphorylation by PKN1 inhibits the formation of filaments. Phosphorylated at Ser-56 by CDK5 during neutrophil secretion in the cytoplasm. Phosphorylated by STK33.
    O-glycosylated during cytokinesis at sites identical or close to phosphorylation sites, this interferes with the phosphorylation status.
    S-nitrosylation is induced by interferon-gamma and oxidatively-modified low-densitity lipoprotein (LDL(ox)) possibly implicating the iNOS-S100A8/9 transnitrosylase complex.
  • 細胞内局在Cytoplasm.
  • Information by UniProt
  • 参照データベース
  • 製品の状態Vimentin is found in connective tissue and in the cytoskeleton.
  • 別名
    • CTRCT30 antibody
    • Epididymis luminal protein 113 antibody
    • FLJ36605 antibody
    • HEL113 antibody
    • VIM antibody
    • VIME_HUMAN antibody
    • Vimentin antibody
    see all

Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker 画像

  • Anti-vimentin (ab92547) staining in E17 rat cheek sections using immunohistochemistry (formaldehyde-fixed, paraffin-embedded sections). Heat-mediated antigen retrieval was carried out using citric acid. Samples were incubated with primary antibody (1/2000) for two hours at room temperature. A biotin-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.

    Image courtesy of Mr Carl Hobbs, Kings College London.

  • Anti-vimentin (ab92547) staining in adult mouse brain (the dentate gyrus region of the hippocampus) using immunohistochemistry (formaldehyde-fixed, paraffin-embedded sections). Heat-mediated antigen retrieval was carried out using citric acid. Samples were incubated with primary antibody (1/2000) for two hours at room temperature. A biotin-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.

    Image courtesy of Mr Carl Hobbs, Kings College London.

  • Anti-vimentin (ab92547) staining in human ovarian cancer tissue using immunohistochemistry (formaldehyde-fixed, paraffin-embedded sections). Heat-mediated antigen retrieval was carried out using citric acid. Samples were incubated with primary antibody (1/2000) for two hours at room temperature. A biotin-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.

    Image courtesy of Mr Carl Hobbs, Kings College London.

  • All lanes : Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab92547) at 1/5000 dilution (purified)

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
    Lane 2 : HEK293 (Human epithelial cell line from embryonic kidney) cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size : 54 kDa
    Observed band size : 54 kDa

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • IHC image of unpurified ab92547 staining Vimentin in human breast adenocarcinoma formalin-fixed paraffin-embedded tissue sections*, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab92547, 1/200 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the negative control (shown on the inset).

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • ab92547 staining Vimentin in HeLa cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab92547 at 5μg/ml and ab7291 at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an Alexa Fluor® 488 Goat anti-Rabbit secondary (ab150081) at 2 μg/ml (shown in green) and Alexa Fluor® 594 Goat anti-Mouse secondary (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.

    Negative controls: 1– Rabbit primary antibody and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.

  • Overlay histogram showing HeLa cells stained with unpurified ab92547 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 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 (ab92547 , 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (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 HeLa cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Triton X-100 used under the same conditions.

  • All lanes : Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab92547) at 1/5000 dilution (purified)

    Lane 1 : mouse brain lysate
    Lane 2 : rat rain lysate

    Secondary
    HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size : 54 kDa
    Observed band size : 54 kDa

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • Immunohistochemical staining of paraffin embedded human cervical carcinoma with purified ab92547 at a working dilution of 1/250. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
  • Immunofluorescence staining of HeLa cells with purified ab92547 at a working dilution of 1/250, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab92547 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

  • Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab92547) at 1/20000 dilution (purified) + COS-1(African green monkey kidney fibroblast-like cell line) cell lysate at 20 µg

    Secondary
    HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size : 54 kDa
    Observed band size : 54 kDa

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • Unpurified ab92547 staining Vimentin in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab92547 at a working concentration of 5μg/ml and ab195889, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor® 594, shown in red) at 1/250 overnight at +4°C, followed by a further incubation at room temperature for 1h with an anti-rabbit AlexaFluor® 488 (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Immunohistochemical staining of paraffin embedded mouse kidney with purified ab92547 at a working dilution of 1/250. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
  • All lanes : Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab92547) at 1/1000 dilution (unpurified)

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) Whole Cell Lysate
    Lane 2 : HEK293 (Human epithelial cell line from embryonic kidney) Whole Cell Lysate
    Lane 3 : Jurkat (Human T cell lymphoblast-like cell line) Whole cell lysate
    Lane 4 : A549 (Human lung carcinoma cell line) Whole cell lysate
    Lane 5 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 6 : PC12 (Rat adrenal gland pheochromocytoma cell line) Whole Cell Lysate
    Lane 7 : HUVEC (Human umbilical vein endothelial cell line) Whole cell lysate
    Lane 8 : A431 (Human epidermoid carcinoma cell line) Whole cell lysate
    Lane 9 : Daudi (Human Burkitt's lymphoma cell line) Whole cell lysate
    Lane 10 : Caco 2 (Human colorectal adenocarcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    IRDye® 800CW Goat Anti-Rabbit at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 54 kDa
    Observed band size : 53 kDa (why is the actual band size different from the predicted?)

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using LI-COR® blocking buffer before being incubated with ab92547 overnight at 4°C. Antibody binding was detected using the IRDye® 800CW Goat Anti-Rabbit secondary at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Odyssey® CLx Imaging System.

  • Immunohistochemical analysis of frozen mouse testis tissue sections labelling Vimentin with ab92547 at a dilution of 1/500. The sections were fixed with Paraformaldehyde and Permeabilized with 0.1% Triton X-100 in PBS. Ab150062 at 1/500 was used as the secondary antibody. Tissue was counterstained with DAPI.

    See Abreview

  • Immunohistochemical analysis of formalin/PFA-fixed paraffin-embedded human cervical carcinoma tissue sections labeling Vimentin with ab92547.

  • Immunohistochemical analysis of formalin/PFA-fixed paraffin-embedded human kidney tissue sections labeling Vimentin with ab92547.

  • Unpurified ab92547 staining vimentin in human Schlemms Canal Endothelium cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with Triton X-100 0.2% and blocked with 10% serum for 30 minutes at 20°C. Samples were incubated with primary antibody (1/200 in DPBS) for 3 hours at 20°C. An undiluted Alexa Fluor®488-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody.

  • Overlay histogram showing HeLa cells fixed in 2% PFA and stained with purified ab92547 at a dilution of 1 in 50 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).
  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD
  • ab92547 staining Vimentin in rat skin tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with 10% buffered normal formalin and blocked with 5% serum for 60 minutes at 21°C; antigen retrieval was by heat mediation in a 10mM Sodium citrate buffer. Samples were incubated with primary antibody (1/200 in blocking buffer) for 12 hours at 4°C. A Cy3®-conjugated donkey anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody.

    See Abreview

  • Fluorescent immunohistochemical analysis of paraffin-embedded human normal kidney tissue using unpurified ab92547. Green- Vimentin red-PI.

  • Fluorescent immunohistochemical analysis of paraffin-embedded human normal colon tissue using unpurified ab92547. Green- Vimentin red-PI

Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab92547) 使用論文

This product has been referenced in:
  • Jia X  et al. Large cervicothoracic myxoinflammatory fibroblastic sarcoma with brachial plexus invasion: A case report and literature review. Oncol Lett 12:1717-1720 (2016). IHC-P ; Human . Read more (PubMed: 27588121) »
  • Su B  et al. Diallyl disulfide suppresses epithelial-mesenchymal transition, invasion and proliferation by downregulation of LIMK1 in gastric cancer. Oncotarget 7:10498-512 (2016). Read more (PubMed: 26871290) »

See all 109 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Flow Cytometry
Sample Human Cell (MDA-MB-231)
Specification MDA-MB-231
Preparation Cell harvesting/tissue preparation method: Cell dissociation buffer
Sample buffer: Enzyme free
Fixation Paraformaldehyde
Permeabilization Yes - 70% Methanol
Gating Strategy Live
Username

Abcam user community

Verified customer

投稿 Feb 18 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: RT°C
Sample Mouse Cell (NIH/3T3)
Specification NIH/3T3
Permeabilization Yes - 0.2% Triton-X-100
Fixative Formaldehyde
Username

Abcam user community

Verified customer

投稿 Aug 12 2013

Thank you for your enquiry. These products are sold as tissue culture supernatant. Unpurified antibodies, such as those sold as whole antiserum, ascites or tissue culture supernatant generally will not have a concentration stated on the datasheet be...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Rhesus monkey Tissue sections (Brain)
Specification Brain
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH6.0
Permeabilization No
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 20% · Temperature: 37°C
Username

Mr. Peter Sullivan

Verified customer

投稿 Nov 16 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Schlemms Canal Endothelium)
Specification Schlemms Canal Endothelium
Fixative Formaldehyde
Permeabilization Yes - 0.2% Triton X-100
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 20°C
Username

Dr. Thomas Read

Verified customer

投稿 Oct 31 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Rat Tissue sections (Skin)
Specification Skin
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrated Buffer, pH6.0
Permeabilization No
Blocking step BSA as blocking agent for 25 minute(s) · Concentration: 1% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Jul 22 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Liver - (after bile duct ligation surgery))
Specification Liver - (after bile duct ligation surgery)
Fixative Paraformaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate buffer pH6.0
Permeabilization No
Blocking step Serum as blocking agent for 20 minute(s) · Concentration: 20% · Temperature: 21°C
Username

Abcam user community

Verified customer

投稿 Jun 20 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Rat Tissue sections (Liver tissue)
Specification Liver tissue
Fixative Paraformaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate buffer pH6.0
Permeabilization No
Blocking step Serum as blocking agent for 20 minute(s) · Concentration: 20% · Temperature: 21°C
Username

Dr. Philip Probert

Verified customer

投稿 Jun 20 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (AD Brain)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Specification AD Brain
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

投稿 Oct 24 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Brain)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Specification Brain
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

投稿 Oct 24 2016

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"