Anti-Vimentin (phospho S55) antibody [4A4] (ab22651)

Overview

  • Product nameAnti-Vimentin (phospho S55) antibody [4A4]
    See all Vimentin primary antibodies
  • Description
    Mouse monoclonal [4A4] to Vimentin (phospho S55)
  • SpecificityDoes not detect non-phosphorylated Vimentin or Vimentin phosphorylated by camp dependant kinase, protein kinase C or Ca2+ calmodulin dependant kinase II.
  • Tested applicationsSuitable for: WB, ELISA, ICC/IF, IHC-P, IHC-Fr, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Ferret
  • Immunogen

    Synthetic peptide:

    SLYSSSPGGAYV

    corresponding to amino acids 50 - 61 of Mouse Vimentin, containing phosphorlyated serine 55, and conjugated to KLH.

  • Positive control
    • The antibody was tested positive with the following cell lines via WB: U251, NIH-3T3, and 3Y1-B.

Properties

Applications

Our Abpromise guarantee covers the use of ab22651 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
WB Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 57 kDa (predicted molecular weight: 54 kDa).
ELISA Use at an assay dependent concentration.
ICC/IF Use a concentration of 1 µg/ml.
IHC-P Use at an assay dependent concentration. PubMed: 20436478
IHC-Fr Use at an assay dependent concentration. PubMed: 20436478
Flow Cyt Use 1µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

Target

  • 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.
  • Post-translational
    modifications
    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.
  • Cellular localizationCytoplasm.
  • Information by UniProt
  • Database links
  • FormVimentin is found in connective tissue and in the cytoskeleton.
  • Alternative names
    • CTRCT30 antibody
    • Epididymis luminal protein 113 antibody
    • FLJ36605 antibody
    • HEL113 antibody
    • VIM antibody
    • VIME_HUMAN antibody
    • Vimentin antibody
    see all

Anti-Vimentin (phospho S55) antibody [4A4] images

  • ab22651 at 1µg/ml staining Vimentin (phospho S55) in human U251 cells by Immunocytochemistry/ Immunofluorescence.

  • Lane 1 :
    Lane 2 :

    Lane 1 : U251 interphase cells
    Lane 2 : U251 M phase cells

    Lysates/proteins at 5 µg/ml per lane.


    Predicted band size : 54 kDa

    Western Blot analysis using ab22651

  • Overlay histogram showing HeLa cells stained with ab22651 (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 (ab22651, 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 IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

References for Anti-Vimentin (phospho S55) antibody [4A4] (ab22651)

This product has been referenced in:
  • Wong FK  et al. Sustained Pax6 Expression Generates Primate-like Basal Radial Glia in Developing Mouse Neocortex. PLoS Biol 13:e1002217 (2015). IF . Read more (PubMed: 26252244) »
  • Ranchoux B  et al. Endothelial-to-mesenchymal transition in pulmonary hypertension. Circulation 131:1006-18 (2015). WB, IHC-Fr ; Human . Read more (PubMed: 25593290) »

See all 10 Publications for this product

Product Wall

While I am not able to replace this product as it is outside of the Abpromise period, I would be happy to provide a discount of 30% off purchase of one vial of ab59408 by using the code XXXXX.

Thank you for your reply.

I would be happy to make the exception this time for you and replace ab22651 with another primary antibody. Could you please provide the Abcam order conformation number or the PO and date of purchase along with th...

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Thank you for providing an Abreview for ab22651 anti-Vimentin antibody.

I am sorry to see that this product did not perform as expected in ICC/IF on human colon carcinoma samples. We guarantee our products to work as stated on the datashee...

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Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Colon carcinoma)
Specification Colon carcinoma
Fixative Paraformaldehyde
Permeabilization Yes - 0.5% Triton in PBS
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 37°C
Username

Dr. Eleni Petsalaki

Verified customer

Submitted Dec 04 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Ferret Tissue sections (Brain)
Specification Brain
Fixative Paraformaldehyde
Permeabilization Yes - Triton-X
Blocking step 0.1 M glycine buffer as blocking agent for 30 minute(s) · Concentration: 7.5µg/mL · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Aug 31 2010

Thank you for your enquiry. Mouse vimentin is Ser-55, which is Ser-56 in human vimentin. The antibody will recognize either (the immunogen is from mouse vimentin so the mouse phosphorylation site was chosen for the product name/description). I ...

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Thank you for your phone call. This antibody was tested positive with the following cell lines via WB: U251, NIH-3T3, and 3Y1-B. I would recommend using one of these as a positive control. It also tests positive with the phospho-Vimentin (Ser55) pepti...

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