Anti-Vimentin抗体[V9] - Cytoskeleton Marker (ab8069)

概述

性能

应用

Our Abpromise guarantee covers the use of ab8069 in the following tested applications.

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

应用 Ab评论 说明
ICC/IF Use a concentration of 5 µg/ml.
Flow Cyt Use 0.1-1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

IHC-FoFr Use at an assay dependent concentration.
IHC-P Use a concentration of 0.5 - 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-Fr 1/300.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 57 kDa (predicted molecular weight: 54 kDa).

靶标

  • 功能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.
  • 组织特异性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.
  • 疾病相关Cataract 30
  • 序列相似性Belongs to the intermediate filament family.
  • 结构域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.
  • 翻译后修饰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.
  • 细胞定位Cytoplasm.
  • Information by UniProt
  • 数据库链接
  • 形式Vimentin is found in connective tissue and in the cytoskeleton.
  • 别名
    • CTRCT30 antibody
    • Epididymis luminal protein 113 antibody
    • FLJ36605 antibody
    • HEL113 antibody
    • VIM antibody
    • VIME_HUMAN antibody
    • Vimentin antibody
    see all

Anti-Vimentin antibody [V9] - Cytoskeleton Marker 图像

  • IHC image of ab8069 staining Vimentin in normal human kidney 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 ab8069, 1/500 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

  • ab8069 staining Vimentin in HeLa cells. The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab8069 at 1/100 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at 2µg/ml (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • All lanes : Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) Whole Cell Lysate
    Lane 2 : MOLT4 (Human lymphoblastic leukemia cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 54 kDa
    Observed band size : 57 kDa (why is the actual band size different from the predicted?)


    Exposure time : 20 minutes
  • Overlay histogram showing SV40LT-SMC cells stained with ab8069 (red line). The cells were fixed with 4% formaldehyde (10 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 (ab8069, 0.1μg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

    This antibody gave a positive signal in SV40LT-SMC cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Triton X-100 for 20 min used under the same conditions.

  • ab8069 staining Vimentin in Human oral cavity tissue sections by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).

    Tissue was fixed with paraformaldehyde and blocked with ab64226 Protein Block for 5 minutes at 25°C; antigen retrieval was by heat mediation in pH 6 buffer . Samples were incubated with primary antibody (1/1000 in 10% NGS) for 16 hours at 4°C. An undiluted biotinylated goat anti-rabbit polyclonal IgG was used as the secondary antibody.

    See Abreview

  • IHC image of Vimentin staining in human Hodgkin's Lymphoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. 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 ab8069, 1µg/ml, 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
  • ab8069 staining Vimentin in human epithelial ovarian serous adenocarcinoma cell line SKOV-3 by Immunocytochemistry/ Immunofluorescence.
    Samples were fixed with 4% PFA in PBS pH 7.4 and then permeabilised using 0.2% saponin for 30 minutes. A blocking step was performed using 1% BSA/PBS for 1 hour. Samples were then incubated with ab8069 at a 1/200 dilution in 1% BSA/PBS for 1 hour. The secondary antibody was a goat anti-mouse Alexa 488 (green) diluted 1/1000, 1% BSA/PBS for 1 hour. Samples were then incubated with phalloidin (red for actin staining) in 1% BSA/PBS for 45 minutes and counterstained with DAPI (blue for nuclei staining) in PBS for 45 minutes.
  • ab8069 staining Vimentin in pig cardiac fibroblasts by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with acetone and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody (1/100 in PBS pH 7.4) for 30 minutes. A FITC-conjugated goat anti-mouse IgG polyclonal (1/20) was used as the secondary antibody.

    See Abreview

  • ab8069 staining Vimentin in rat glial cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde and blocked with 1% BSA for 10 minutes at 25°C. Samples were incubated with primary antibody (1/200) for 10 hours at 4°C. An Alexa Fluor® 555-conjugated anti-mouse IgG polyclonal (1/300) was used as the secondary antibody.

    See Abreview

  • ICC/IF image of ab8069 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab8069, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1h.Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Overlay histogram showing MDA-MB-231 cells stained with ab8069 (red line). The cells were fixed with 4% formaldehyde (10 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 (ab8069, 1μg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

    This antibody gave a positive signal in MDA-MB-231 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Triton X-100 for 20 min used under the same conditions.

Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069)参考文献

This product has been referenced in:
  • Salzman DW  et al. miR-34 activity is modulated through 5'-end phosphorylation in response to DNA damage. Nat Commun 7:10954 (2016). IP, WB ; Human . Read more (PubMed: 26996824) »
  • Ren M  et al. A biofidelic 3D culture model to study the development of brain cellular systems. Sci Rep 6:24953 (2016). ICC/IF ; Rat . Read more (PubMed: 27112667) »

See all 33 Publications for this product

Product Wall

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 (Kidney)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate buffer (pH 6)
Permeabilization No
Specification Kidney
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Fixative Paraformaldehyde
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提交于 Aug 02 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Dog Tissue sections (canine soft tissue sarcoma)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9
Permeabilization No
Specification canine soft tissue sarcoma
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: 20°C
Fixative Formaldehyde
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提交于 Dec 18 2015

Application Western blot
Loading amount 10 µg
Gel Running Conditions Reduced Denaturing (10%)
Sample Pig Tissue lysate - whole (Heart)
Specification Heart
Treatment 5 pig are treated with pacemaker for 14 days (pm)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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提交于 Jan 21 2015

Application IHC - Wholemount
Sample Human Tissue (Brain)
Specification Brain
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提交于 Jan 06 2015

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (12% Gel)
Sample Rat Cell lysate - whole cell (mixed glial cells)
Specification mixed glial cells
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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提交于 Dec 05 2014

Application Immunocytochemistry/ Immunofluorescence
Sample Rat Cell (Glial Cells)
Specification Glial Cells
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative Formaldehyde
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提交于 Jul 14 2014

Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: CItrate Buffer
Sample Rat Tissue sections (brain)
Specification brain
Blocking step Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative Formaldehyde
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Dr. Yasir Ahmed Syed

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提交于 Jul 09 2014

Application Western blot
Loading amount 10 µg
Gel Running Conditions Reduced Denaturing
Sample Human Tissue lysate - whole (Heart)
Specification Heart
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C
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提交于 Jan 22 2014

Application Immunocytochemistry/ Immunofluorescence
Sample Pig Cell (cardiac fibroblasts)
Specification cardiac fibroblasts
Permeabilization Yes - Triton X-100 0.1%
Fixative Acetone
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提交于 Jan 09 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (oral squamous cell carcinoma)
Specification oral squamous cell carcinoma
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative Paraformaldehyde
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提交于 Sep 10 2013

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