Product nameAnti-Vimentin (phospho S56) antibody [EPR21084]
See all Vimentin primary antibodies
DescriptionRabbit monoclonal [EPR21084] to Vimentin (phospho S56)
Tested applicationsSuitable for: WB, Dot blot, ICC/IF, IP, Flow Cytmore details
Species reactivityReacts with: Human
Synthetic peptide within Human Vimentin aa 1-100 (phospho S56). The exact sequence is proprietary.
Database link: P08670
Storage instructionsShipped 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 bufferPreservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
PurityProtein A purified
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.
|WB||1/1000. Detects a band of approximately 57 kDa (predicted molecular weight: 54 kDa).|
|Flow Cyt||Use at an assay dependent concentration.|
FunctionVimentins 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 specificityHighly 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 diseaseCataract 30
Sequence similaritiesBelongs to the intermediate filament family.
DomainThe 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.
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.
- Information by UniProt
FormVimentin is found in connective tissue and in the cytoskeleton.
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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.
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 seconds
Blocking/Dilution buffer: 5% NFDM/TBST.
Ab217673 staining Vimentin (phospho S56) in HeLa (human cervix adenocarcinoma epithelial cell) cells by 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)
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.
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.
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.
ab217673 has not yet been referenced specifically in any publications.