使用敲除细胞株进行验证

Anti-Vimentin抗体[VI-10] (ab20346)

概述

  • 产品名称
    Anti-Vimentin抗体[VI-10]
    参阅全部 Vimentin 一抗
  • 描述
    小鼠单克隆抗体[VI-10] to Vimentin
  • 特异性
    The antibody VI-10 reacts with vimentin, a 57 kDa intermediate filament expressed in variety of mesenchymal and mesodermal cell types.
  • 经测试应用
    适用于: ICC/IF, IP, WB, IHC-P, Flow Cytmore details
  • 种属反应性
    与反应: Mouse, Rat, Chicken, Dog, Human, Pig
  • 免疫原

    Full length native protein (purified) corresponding to Vimentin.

  • 阳性对照

性能

应用

Our Abpromise guarantee covers the use of ab20346 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 1 µg/ml.
IP Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 54 kDa.
IHC-P Use a concentration of 1 - 5 µg/ml.
Flow Cyt Use 1µg for 106 cells.

ab91545 - Mouse monoclonal IgM, is suitable for use as an isotype control with this antibody.

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 [VI-10] 图像

  • ab20346 staining Vimentin in wild-type HAP1 cells (top panel) and Vimentin 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 ab20346 at 1μg/ml dilution and ab202272 at 1/250 dilution (shown in pseudo-color red) overnight at +4°C. The cells were then incubated with ab150117 (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488)) at 1/1000 dilution for 1 hour. Nuclear DNA was labelled in blue with DAPI.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • ab20346 at a 1/500 dilution staining mouse dissociated neural precursor cells by immunocytochemistry. The antibody was incubated with the cells for 1 hour and then detected using a goat anti-mouse Alexa-Fluor 568 (red stain) ab175473>ab175473). The image also shows a blue nuclear counterstain. Confocal image is shown with 2x zoom detail in top right corner.

    This image is courtesy of an Abreview submitted by Randal Moldich on 22 February 2006.

    See Abreview

  • Human normal skin. Staining is localised to cytoplasm. Left panel: with primary antibody at 1 ug/mL. Right panel: isotype control. Sections were stained using an automated system DAKO Autostainer Plus , at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers citrate pH6.1 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 mouse 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 we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

  • Immunocytochemistry/ Immunofluorescence analysis of chicken postnatal erythrocytes labelling Vimentin (green) with ab20346. Tubulin was counterstained red with anti-alpha-tubulin and DAPI was used as a nuclear counterstain. 


  • Predicted band size : 54 kDa

    Image from Gao MQ et al, J Cell Sci. 2010 Oct 15;123(Pt 20):3507-14. Epub 2010 Sep 14, Fig 3. DOI 10.1242/?jcs.072900

    ab20346 used at a 1/1000 dilution in Western Blot.5-20µg of total protein from each sample was loaded.

  • Overlay histogram showing NIH3T3 cells stained with ab20346 (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 (ab20346, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was a goat anti-mouse DyLight® 488 (IgG; H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgM [ICIGM] (ab91545, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in NIH3T3 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Triton used under the same conditions.

  • Immunofluorescence staining of RBL rat basophiilioc cell line with anti-Vimentin (VI--10). Nuclei are stained with DAPI (blue).

     

  •  ab20346 at a 1/1000 dilution staining rat astrocyte cells by immunocytochemistry. The antibody was incubated with the cells for 1 hour and then detected using a goat anti-mouse Alexa-Fluor ® 568 (ab175473).

    This image is courtesy of an Abreview submitted by Randal Moldrich on 8 February 2006

    See Abreview

Anti-Vimentin antibody [VI-10] (ab20346)参考文献

This product has been referenced in:
  • van Marion MH  et al. Behavior of CMPCs in unidirectional constrained and stress-free 3D hydrogels. J Mol Cell Cardiol 87:79-91 (2015). IF ; Human . Read more (PubMed: 26278995) »
  • Zerr P  et al. Vitamin D receptor regulates TGF-ß signalling in systemic sclerosis. Ann Rheum Dis N/A:N/A (2014). Human . Read more (PubMed: 24448349) »

See all 25 Publications for this product

Product Wall

Application
Immunocytochemistry
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1.5% · Temperature: 25°C
Sample
Mouse Cultured Cells (thoracic aortic primary smooth muscle cells)
Specification
thoracic aortic primary smooth muscle cells
Permeabilization
Yes - 0.25% TritonX-100
Fixative
Paraformaldehyde
Username

Mr. Mc Shen

Verified customer

提交于 Dec 15 2014

Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (12,5)
Sample
Human Tissue lysate - whole (human Urether)
Specification
human Urether
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Nov 11 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Flow Cytometry
Fixation
Frozen no other fixative
Permeabilization
Yes - 0.1% Triton-X
Sample
Human Cell (Brain)
Specification
Brain
Gating Strategy
forward scatter/side scatter (all cells)
Preparation
Cell harvesting/tissue preparation method: Homogenized frozen tissue
Sample buffer: Tris/EDTA/sucrose/Triton
Username

Abcam user community

Verified customer

提交于 Sep 06 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 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C
Sample
Human Cell (mammalian cancer cells)
Specification
mammalian cancer cells
Permeabilization
Yes - Tween20
Fixative
Formaldehyde
Username

Mr. Gabriel Ortega

Verified customer

提交于 Jul 01 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: 1% · Temperature: 37°C
Sample
Human Cell (Colon carcinoma)
Specification
Colon carcinoma
Permeabilization
Yes - 0.5% Triton in PBS
Fixative
Paraformaldehyde
Username

Dr. Eleni Petsalaki

Verified customer

提交于 Dec 04 2012

Thank you for contacting us. I am sorry to hear that the suggestions did not improve the performance of this product.

Because we carry over 70,000 products, it isn't feasible for us to keep small sample sizes of our products.

Ho...

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Thank you for taking time to contact Abcam. I am sorry to hear that this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (mouse cardiac fibroblasts)
Loading amount
20 µg
Specification
mouse cardiac fibroblasts
Gel Running Conditions
Reduced Denaturing (10)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Apr 02 2012

Thank you for contacting us.

The protocol we have for GMA staining is as follows;

Place biopsy immediately in ice cold acetone containing protease inhibitors
Fix overnight -20°C
Replace fixative with acetone (room tem...

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Thank you for your enquiry. I have contacted the originator of this antibody, and they have kindly provided the following information regarding the IP testing: 'The lysis was done by 1% lauryl maltoside + protease inhibitors. I am sorry we do not...

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

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