Recombinant Anti-Vimentin (phospho S56) antibody [EPR21084] (ab217673)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR21084] to Vimentin (phospho S56)
- Suitable for: WB, Dot blot, ICC/IF, IP, Flow Cyt (Intra)
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Vimentin (phospho S56) antibody [EPR21084]
See all Vimentin primary antibodies -
Description
Rabbit monoclonal [EPR21084] to Vimentin (phospho S56) -
Host species
Rabbit -
Tested applications
Suitable for: WB, Dot blot, ICC/IF, IP, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR21084 -
Isotype
IgG -
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 ab217673 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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WB |
1/1000. Detects a band of approximately 57 kDa (predicted molecular weight: 54 kDa).
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Dot blot |
1/1000.
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ICC/IF |
1/1000.
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IP |
1/30.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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Notes |
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WB
1/1000. Detects a band of approximately 57 kDa (predicted molecular weight: 54 kDa). |
Dot blot
1/1000. |
ICC/IF
1/1000. |
IP
1/30. |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
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
- Omim: 193060 Human
- SwissProt: P08670 Human
- Unigene: 455493 Human
- Unigene: 691131 Human
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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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All lanes : Anti-Vimentin (phospho S56) antibody [EPR21084] (ab217673) at 1/1000 dilution
Lane 1 : Untreated HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : HeLa (human epithelial cell line from cervix adenocarcinoma) treated with 100 ng/ml nocodazole (ab120630) for 18 hours, whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Developed using the ECL technique.
Predicted band size: 54 kDa
Observed band size: 57 kDa why is the actual band size different from the predicted?
Exposure time: 23 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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Ab217673 staining Vimentin (phospho S56) in HeLa (human cervix adenocarcinoma epithelial cell) cells by intracellular flow cytometry. Cells were fixed using 80% Methanol and permeabilized with 0.1% Tween-20. The sample was incubated with primary antibody at 1/500 dilution. An Alexa Fluor®488 Goat anti-rabbit IgG (ab150077) was used as a secondary antibody. Rabbit monoclonal IgG (ab172730) was used as an isotype control (left). Cells were pre-treated with 20 µg/ml RNase A for 30 minutes to eliminate the non-specific binding between RNA and PI (Propidium iodide). Vimentiin (phospho S56) is highly expressed in mitotic cells. (PMID: 16260496)
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Dot blot analysis of Vimentin (phospho S56) labeled with ab217673 at 1/1000 dilution.
Lane 1: Vimentin (phospho S56) peptide.
Lane 2: Vimentin non-phospho peptide.Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution was used as secondary antibody.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 1 minute.
The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling Vimentin (phospho S56) with ab217673 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing strong positive staining in HeLa cells in M phase.
The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).
Secondary antibody only control: PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
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Vimentin (phospho S56) was immunoprecipitated from 0.35 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) treated with 100 ng/ml nocodazole (ab120630) for 18 hours, whole cell lysate with ab217673 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab217673 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: HeLa treated with 100 ng/ml nocodazole (ab120630) for 18 hours, whole cell lysate 10 μg (Input).
Lane 2: ab217673 IP in HeLa treated with 100 ng/ml nocodazole (ab120630) for 18 hours, whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab217673 in HeLa treated with 100 ng/ml nocodazole (ab120630) for 18 hours, whole cell lysate.Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (8)
ab217673 has been referenced in 8 publications.
- Zhou H et al. miR-1266-3p Suppresses Epithelial-Mesenchymal Transition in Colon Cancer by Targeting P4HA3. Anal Cell Pathol (Amst) 2022:1542117 (2022). PubMed: 35433237
- Tan Q et al. VEGF-B targeting by aryl hydrocarbon receptor mediates the migration and invasion of choriocarcinoma stem-like cells. Cancer Cell Int 22:221 (2022). PubMed: 35773697
- Zhang S & Chen R LINC01140 regulates osteosarcoma proliferation and invasion by targeting the miR-139-5p/HOXA9 axis. Biochem Biophys Rep 31:101301 (2022). PubMed: 35800618
- Li Y et al. Upregulation of SIX4 indicates poor clinical outcome and promotes tumor growth and cell metastasis in esophageal squamous cell carcinoma. Thorac Cancer 12:752-759 (2021). PubMed: 33481352
- Tian C et al. Linc-ROR facilitates progression and angiogenesis of hepatocellular carcinoma by modulating DEPDC1 expression. Cell Death Dis 12:1047 (2021). PubMed: 34741030
- Sun L et al. MicroRNA-615 functions as a tumor suppressor in osteosarcoma through the suppression of HK2. Oncol Lett 20:226 (2020). PubMed: 32968448
- Chen W et al. TERF1 downregulation promotes the migration and invasion of the PC3 prostate cancer cell line as a target of miR-155. Mol Med Rep 22:5209-5218 (2020). PubMed: 33174061
- Liu J et al. Bnip3 interacts with vimentin, an intermediate filament protein, and regulates autophagy of hepatic stellate cells. Aging (Albany NY) 13:957-972 (2020). PubMed: 33290258