使用敲除细胞株进行验证

Anti-Vimentin抗体[RV202] (ab8978)

概述

  • 产品名称
    Anti-Vimentin抗体[RV202]
    参阅全部 Vimentin 一抗
  • 描述
    小鼠单克隆抗体[RV202] to Vimentin
  • 特异性
    This antibody reacts exclusively with vimentin
  • 经测试应用
    适用于: Flow Cyt, ICC, IHC-Fr, WB, IHC-FoFr, IP, IHC-P, ICC/IF, IHC (PFA fixed)more details
  • 种属反应性
    与反应: Mouse, Rat, Goat, Chicken, Hamster, Cow, Dog, Human, Xenopus laevis, Monkey, Zebrafish
  • 免疫原

    Fusion protein corresponding to Bovine Vimentin. RV202 is a mouse monoclonal IgG1 antibody derived by fusion of SP2/0-Ag14 mouse myeloma cells with spleen cells from a BALB/c mouse immunized with a vimentin extract of bovine lens.

  • 阳性对照

性能

应用

Our Abpromise guarantee covers the use of ab8978 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
Flow Cyt 1/100 - 1/200.

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

ICC 1/20.
IHC-Fr Use at an assay dependent concentration. Recommended range is 1/100 - 1/200 for Immunohistochemistry with avidin-biotinylated horseradish peroxidase complex (ABC) as detection. For PFA fixed tissue use at 1/1000.
WB 1/100 - 1/1000. Detects a band of approximately 57 kDa.
IHC-FoFr 1/500.
IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
ICC/IF Use a concentration of 1 µg/ml.
IHC (PFA fixed) 1/1000.

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

图片

  • ab8978 staining Vimentin in wild-type HAP1 cells (top panel) and VIM knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), 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 with ab8978 at 1μg/ml and ab202272 (Rabbit monoclonal [EP1332Y] to alpha Tubulin (Alexa Fluor® 594)) at 1/250 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with ab150117 (Goat secondary antibody to Mouse IgG (Alexa Fluor® 488)) 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).

  • ab8978 were fixed with paraformaldehyde, permeabilized with PBS and 0.5% Triton ×100 and blocking with 0.1% BSA + 10% Goat Serum at 250C for 30 minutes was performed. Samples were incubated with primary antibody (1/250: in PBS, 0.1% BSA and 10% Goat Serum) for 12 hours at 4°C. An Alexa Fluor®594-conjugated goat polyclonal to mouse IgG was used undiluted as secondary antibody.

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  • ab8978 staining Vimentin in Human fetal kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with CAS-Block for 1 hour at 25°C; antigen retrieval was by heat mediation using OmniPrep (pH 9). Samples were incubated with primary antibody (1/500) for 1 hour at 25°C. An Alexa Fluor® 555-conjugated Donkey polyclonal (1/200) was used as the secondary antibody.

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  • Immunofluorescence staining images of 9 day old zebrafish embryos.
    ab8978 reacts with in connective tissue cells and bloodvessels. Frozen sample treated with Acetone:Methanol 1:1, antibody diluted 1/100 and incubated for 45 minutes at room temperature.

  • Overlay histogram showing HeLa cells stained with ab8978 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Triton 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 (ab8978, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This anti-Vimentin antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Triton used under the same conditions.

  •   ab8978 staining Vimentin - Neural Stem Cell Marker in Human Colon fibroblasts by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with methanol, permeabilized in 0.1% Triton and blocked with 0.25% serum free protein blocker for 20 minutes at 28°C. Samples were incubated with primary antibody (1/100 in antibody diluent) for 2 hours at 28°C. ab6785 Goat polyclonal anti-Mouse IgG - H&L (FITC) (1/800) was used as the secondary antibody. Nuclei were counterstained with propidium iodide.

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  • ab8978 staining Vimentin in Dog soft tissue sarcoma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 15% serum for 1 hour at 20°C; antigen retrieval was by heat mediation in a Tris/EDTA pH9 buffer. Samples were incubated with primary antibody (1/100 in TBS) for 18 hours at 20°C. A Alexa Fluor® 647-conjugated Goat anti-mouse IgG polyclonal (1/400) was used as the secondary antibody.

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  • IHC-Fr image of Ed18 rat stained with ab8978. Fresh frozen sections  were  incubated in  10% normal donkey serum in 0.1% PBS- and 0.3% triton X100  for 1h to permeabilise the tissues and block non-specific protein-protein interactions. The sectons were then incubated with the ab8978 (1µg/ml) and ferroportin overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 donkey anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 568 (red) donkey anti-mouse at a 1/1000 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. Vimentin expressed in the gut muscles.

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  • IHC-FoFr image of vimentin staining on rat injured cortical sections using ab8978 (1:500). The brain was perfusion fixed using 4% PFA and the sections were permeabilized using 0.1% TritonX in 0.1% PBS. The sections were then blocked using 10% donkey serum for 1 hour at 24°C. ab8978 was diluted 1:500 and incubated with sections for 24 hours using 4°C. The secondary antibody used was donkey polyclonal to rabbit IgG conjugated to Alexa Fluor 488.

  • ab8978 vimentin staining of a tonsilar lymphoma. Note that the epithelium (at the left) is negative.

  •  ab8978 staining Vimentin in bovine chromospheres by ICC/IF (immunocytochemistry/immunofluorescence). Cells were PFA fixed and permeabilized in 0.3% Triton X-100. The primary antibody (1/500) was incubated with the sample for 16 hours at 4°C. An Alexa Fluor® 568-conjugated goat anti-mouse IgG polyclonal (1/500) ab175473 was used as the secondary.  

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  • ab8978 vimentin immunofluorescent staining of cultured bovine lens epithelial cells

  •  ab8978 staining vimentin in human pancreatic adenocarcinoma cells by immunocytochemistry/ immunofluorescence. Cells were PFA fixed and permeabilized in 0.2% Triton X prior to blocking in 3% BSA for 30 minutes at 24°C. The primary antibody was diluted 1/200 and incubated with the sample for 16 hours at 21°C. Alexa fluor® 488 mouse polyclonal to mouse Ig, diluted 1/300 was used as the secondary antibody.

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  • ab8978staining Vimentin in human lung tissue section by Immunohistochemistry (Frozen sections). Tissue samples were fixed with formaldehyde and blocking with 5% commercially available blocking agent was performed at 370C  for 15 minutes. The sample was incubated with primary antibody (1/250) at 370C for 1 hour. A HRP-conjugated Goat polyclonal to mouse IgG was used as secondary antibody at 1/1000 dilution.

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文献

This product has been referenced in:
  • You W  et al. High-Throughput Screening Identifies miR-451 as a Pleiotropic Modulator That Suppresses Gastric Cancer Metastasis. SLAS Technol 22:136-143 (2017). Read more (PubMed: 27780852) »
  • Meng Q  et al. Mammalian Eps15 homology domain 1 promotes metastasis in non-small cell lung cancer by inducing epithelial-mesenchymal transition. Oncotarget 8:22433-22442 (2017). IHC-P ; Human . Read more (PubMed: 27531895) »

See all 83 Publications for this product

客户评价及客户问答

The exact position of epitope sequence recognized by anti vimentin antibody (clone RV202) is not determined however, the antibody must recognize an evolutionary highly conserved epitope, since it shows a broad spectrum of species cross reactivity (from...

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Thank you for contacting us.

Indeed, a mouse-on mouse staining can be quite tricky; however, it is not impossible. The background will depend on the fixation method ( perfusion fixed tissue vs. normally fixed tissue), on the tissue itself ( ...

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Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Nothobranchius furzeri Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 20 · Temperature: 22°C
Fixative
Formaldehyde
Username

Jolien Van houcke

Verified customer

提交于 Apr 11 2017

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (fibroblast)
Permeabilization
No
Specification
fibroblast
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Aug 19 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (melanoma cell line xenograft)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9
Permeabilization
No
Specification
melanoma cell line xenograft
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: 20°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

提交于 Dec 21 2015

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
Username

Abcam user community

Verified customer

提交于 Dec 18 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (fetal kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: commercial - ZYTOMED's OmniPrep (pH 9)
Permeabilization
Yes - 0.05% tween-20
Specification
fetal kidney
Blocking step
commercial - Invitrogen's CAS-Block as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

提交于 Dec 11 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C
Sample
Human Cell (iPS-derived cardiofibroblast)
Specification
iPS-derived cardiofibroblast
Permeabilization
Yes - 0.2% triton X
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

提交于 Aug 04 2014

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (gel 10%)
Sample
Human Cell lysate - whole cell (breast cancer cell line MDAMB231)
Specification
breast cancer cell line MDAMB231
Blocking step
Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
Username

Abcam user community

Verified customer

提交于 Dec 17 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Sample
Rat Cell (Rat Brain post injury)
Specification
Rat Brain post injury
Permeabilization
Yes - 0.1% TritonX in 0.1% PBS
Fixative
Paraformaldehyde
Username

Dr. Ruma Raha-Chowdhury

Verified customer

提交于 Sep 27 2013

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