製品の概要

  • 製品名
    Anti-Vimentin antibody [RV202]
    Vimentin 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [RV202] to Vimentin
  • 由来種
    Mouse
  • 特異性
    This antibody reacts exclusively with vimentin
  • アプリケーション
    適用あり: Flow Cyt, ICC, IHC-Fr, WB, IHC-FoFr, IP, IHC-P, ICC/IF, IHC (PFA fixed)more details
  • 種交差性
    交差種: 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.

  • ポジティブ・コントロール
  • 特記事項

    Abcam recommended secondaries - Goat Anti-Mouse HRP (ab205719) and Goat Anti-Mouse Alexa Fluor® 488 (ab150113).

    See other anti-mouse secondary antibodies that can be used with this antibody.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

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

画像

  • Silencing of Lamin A/C in unmethylated neuroblastoma cells induces changes in different cytoskeletal components

    Immunofluorescence staining showing changes in SK-N-SH-lamin-A/C-shRNA compared with SK-N-SH-scramble-shRNA in (A) β-actin filaments, (B) F-actin filaments. (C) Vimentin filaments, and (D) α-tubulin. Lamin A/C is shown in green; β-actin, F-actin, vimentin, and α-tubulin are shown in red.DNA is stained in blue (DAPI). Scale bar, 10μM.

    Vimentin is detected using ab8978 at 1/100 dilution. Neuroblastoma cells were fixed with 4% paraformaldehyde and permeabilized using 0.5% Triton X-100.

    (From Figure 5C of Rauschert et al)

  • 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).

  • Paraffin-embedded human colon tissue stained for Vimentin using ab8978 at 1/100 dilution in immunohistochemical analysis.

  • 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.

    See Abreview

  • 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.

  • 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.

  • 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.

    See Abreview

  •   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.

    See Abreview

  • 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.

    See Abreview

  • 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.

    See Abreview

  • 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.  

    See Abreview

  • 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.

    See Abreview

  • 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.

    See Abreview

参考文献

This product has been referenced in:
  • Gao J  et al. In vitro investigation of the mechanism underlying the effect of ginsenoside on the proliferation and differentiation of neural stem cells subjected to oxygen-glucose deprivation/reperfusion. Int J Mol Med 41:353-363 (2018). Read more (PubMed: 29138802) »
  • Yang HJ  et al. GP73 promotes invasion and metastasis of bladder cancer by regulating the epithelial-mesenchymal transition through the TGF-ß1/Smad2 signalling pathway. J Cell Mol Med 22:1650-1665 (2018). WB ; Human . Read more (PubMed: 29349903) »
See all 152 Publications for this product

レビューと Q&A

1-10 of 46 Abreviews or Q&A

Question
Answer

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 human to zebrafish).
For more information on the evolutionary conserved regions in the vimentin protein I would like to refer your customer to the paper by Herrmann et al (1996) Journal of Cell Science 109, 569-578.

Also the antibody does not seem to recognize either the amino- (N-) or carboxy (C-) terminus, since it reacts with the typical vimentin breakdown products that have a more acidic pI than the intact protein and are therefore known to lack about 10 kDa at their N-terminus. Furthermore, a vimentin construct that lacks its C-terminus (amino acids 408-463) is still recognized by the antibody. This information can be found in Krimpenfort et al (1988) EMBO Journal 7, 941-947.

Read More

Answer

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 ( eg. kidneys vs. bone) and the experimental settings.


One way to make sure to decrease or to completelyerase the background is to block the endogenous mouse IgG with a non- conjugated secondary antibody before you incubate with your target antibody. I would suggest to use a Fab or F(ab)2 fragment antibody in this case as they will bound in a higher density than whole IgG antibodies.

You can find the aforementioned antibodies here:


https://www.abcam.com/index.html?datasheet=98754 (or use the following: https://www.abcam.com/index.html?datasheet=98754).

https://www.abcam.com/index.html?datasheet=6668 (or use the following: https://www.abcam.com/index.html?datasheet=6668).



Another way is to use our hassle- free Mouse on Mouse (M.O.M) Polymer IHC Kit, which contains a special rodent block solution:

https://www.abcam.com/index.html?datasheet=127055 (or use the following: https://www.abcam.com/index.html?datasheet=127055).


In addition, we also received an Abreview (17 July 2009) from one of our customers who rated this antibody highly in IHC. You can contact the author via our website if you like.

Read More
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (Synovial tissue)
Antigen retrieval step
Enzymatic - Buffer/Enzyme Used: PBS/Protease
Permeabilization
No
Specification
Synovial tissue
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 20% · Temperature: 20°C
Fixative
Formaldehyde

Abcam user community

Verified customer

投稿 Aug 09 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Cat Tissue sections (kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9.0
Permeabilization
No
Specification
kidney
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: RT°C
Fixative
10% NBF

Abcam user community

Verified customer

投稿 Jun 29 2018

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)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TE pH9
Permeabilization
No
Specification
liver
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% normal buffered formalin

Abcam user community

Verified customer

投稿 Feb 26 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TE pH9
Permeabilization
No
Specification
lung
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% normal buffered formalin

Abcam user community

Verified customer

投稿 Feb 19 2018

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 (stomach tumor)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TE pH9
Permeabilization
No
Specification
stomach tumor
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% normal buffered formalin

Abcam user community

Verified customer

投稿 Feb 19 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (tumor xenograft)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TE pH9
Permeabilization
No
Specification
tumor xenograft
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
10% normal buffered formalin

Abcam user community

Verified customer

投稿 Feb 19 2018

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

Jolien Van houcke

Verified customer

投稿 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

Abcam user community

Verified customer

投稿 Aug 19 2016

1-10 of 46 Abreviews or Q&A

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

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