Anti-Vimentin antibody [VI-RE/1] (ab3974)
Key features and details
- Mouse monoclonal [VI-RE/1] to Vimentin
- Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-P
- Reacts with: Human
- Isotype: IgG1
Related conjugates and formulations
Overview
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Product name
Anti-Vimentin antibody [VI-RE/1]
See all Vimentin primary antibodies -
Description
Mouse monoclonal [VI-RE/1] to Vimentin -
Host species
Mouse -
Specificity
The antibody VI-RE/1 reacts with human vimentin, a 57 kDa intermediate filament protein expressed on a wide variety of mesenchymal and mesodermal cell types. -
Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-Pmore details -
Species reactivity
Reacts with: Human
Predicted to work with: RatDoes not react with: Mouse, Pig -
Immunogen
Recombinant full length protein corresponding to Human Vimentin.
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Positive control
- ICC: HeLa cells. IHC-P: Human bladder and liver tissue. Flow Cyt (Intra): Human ESS-1 and HeLa cells. WB: HeLa whole cell lysate.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.097% Sodium azide
Constituent: PBS -
Concentration information loading...
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Purity
Protein A purified -
Purification notes
Purified from hybridoma culture supernatant. Purity >95% by SDS-PAGE. -
Clonality
Monoclonal -
Clone number
VI-RE/1 -
Isotype
IgG1 -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab3974 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
Use a concentration of 1 - 4 µg/ml.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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ICC/IF |
Use a concentration of 5 - 10 µg/ml.
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WB |
Use a concentration of 1 - 2 µg/ml. Use under non reducing condition. Detects a band of approximately 57 kDa.
Positive control: LEP-19 cell lysate Negative control: 3T3 mouse cell line Sample preparation: Resuspend approx. 50 mil. cells in 1 ml cold Lysis buffer (1% laurylmaltoside in 20 mM Tris/Cl, 100 mM NaCl pH 8.2, 50 mM NaF including Protease inhibitor Cocktail). Incubate 60 min on ice. Centrifuge to remove cell debris. Mix lysate with reducing Laemmli SDS-PAGE sample buffer. Boil for 3 min in water bath. |
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IHC-P | (1) |
1/400.
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Notes |
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Flow Cyt (Intra)
Use a concentration of 1 - 4 µg/ml. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
ICC/IF
Use a concentration of 5 - 10 µg/ml. |
WB
Use a concentration of 1 - 2 µg/ml. Use under non reducing condition. Detects a band of approximately 57 kDa. Positive control: LEP-19 cell lysate Negative control: 3T3 mouse cell line Sample preparation: Resuspend approx. 50 mil. cells in 1 ml cold Lysis buffer (1% laurylmaltoside in 20 mM Tris/Cl, 100 mM NaCl pH 8.2, 50 mM NaF including Protease inhibitor Cocktail). Incubate 60 min on ice. Centrifuge to remove cell debris. Mix lysate with reducing Laemmli SDS-PAGE sample buffer. Boil for 3 min in water bath. |
IHC-P
1/400. |
Target
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Function
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. -
Tissue specificity
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. -
Involvement in disease
Cataract 30 -
Sequence similarities
Belongs to the intermediate filament family. -
Domain
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. -
Post-translational
modificationsFilament 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. -
Cellular localization
Cytoplasm. - Information by UniProt
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Database links
- Entrez Gene: 7431 Human
- Entrez Gene: 81818 Rat
- Omim: 193060 Human
- SwissProt: P08670 Human
- SwissProt: P31000 Rat
- Unigene: 455493 Human
- Unigene: 691131 Human
- Unigene: 2710 Rat
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Form
Vimentin is found in connective tissue and in the cytoskeleton. -
Alternative names
- CTRCT30 antibody
- Epididymis luminal protein 113 antibody
- FLJ36605 antibody
see all
Images
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Flow cytometry (Intracellular) analysis ESS-1 (human endometrial stromal sarcoma cell line) cells labeling Vimentin with ab3974 (GAM-FITC).
Negative control: human lymphocytes.
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Immunohistochemistry staining (paraffin sections) of Vimentin in human liver using mouse monoclonal antibody [VI-RE/1] (ab3974, diluted 1/400), detected with GAM IgG-Alexa Fluor®488 (diluted 1/200; green), cell nuclei stained with PI (1 µg/ml; orange).
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IHC image of Vimentin staining in human bladder formalin fixed paraffin embedded tissue section, performed on a Leica Bond 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 ab3974, 5µg/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.
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.
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ICC image of ab3974 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 (ab3974, 5µg/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.
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Flow Cytometry (Intracellular) analysis of HeLa cells labeling Vimentin with ab3974 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween 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 (ab3974, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. 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.
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Anti-Vimentin antibody [VI-RE/1] (ab3974) at 5 µg/ml + HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
Secondary
Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Observed band size: 57 kDa why is the actual band size different from the predicted?
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (4)
ab3974 has been referenced in 4 publications.
- Li B et al. The MSC-Derived Exosomal lncRNA H19 Promotes Wound Healing in Diabetic Foot Ulcers by Upregulating PTEN via MicroRNA-152-3p. Mol Ther Nucleic Acids 19:814-826 (2020). PubMed: 31958697
- Chen X et al. Down-regulation of microRNA-31-5p inhibits proliferation and invasion of osteosarcoma cells through Wnt/ß-catenin signaling pathway by enhancing AXIN1. Exp Mol Pathol 108:32-41 (2019). PubMed: 30844369
- Hoffmann M et al. Apelin abrogates the stimulatory effects of 17ß-estradiol and insulin-like growth factor-1 on proliferation of epithelial and granulosa ovarian cancer cell lines via crosstalk between APLNR and ERa/IGF1R. Mol Biol Rep 46:6325-6338 (2019). PubMed: 31538301
- Kitayama J et al. CD90+ mesothelial-like cells in peritoneal fluid promote peritoneal metastasis by forming a tumor permissive microenvironment. PLoS One 9:e86516 (2014). Flow Cyt ; Human . PubMed: 24466130