Recombinant
RabMAb

Recombinant Anti-Vimentin (phospho S56) antibody [EPR21084] - BSA and Azide free (ab228854)

Overview

  • Product name

    Anti-Vimentin (phospho S56) antibody [EPR21084] - BSA and Azide free
    See all Vimentin primary antibodies
  • Description

    Rabbit monoclonal [EPR21084] to Vimentin (phospho S56) - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, Dot blot, ICC/IF, IP, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Vimentin aa 1-100 (phospho S56). The exact sequence is proprietary.
    Database link: P08670

  • Positive control

    • ICC/IF: HeLa cells.
  • General notes

    Ab228854 is the carrier-free version of ab217673. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab228854 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab228854 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 at an assay dependent concentration. Detects a band of approximately 57 kDa (predicted molecular weight: 54 kDa).
Dot blot Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

Target

  • 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
    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 localization

    Cytoplasm.
  • Information by UniProt
  • Database links

  • Form

    Vimentin 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

Images

  • 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)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab217673).

  • Vimentin (phospho S56) was immunoprecipitated from 0.35 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) treated with 100 ng/ml nocodazole 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 for 18 hours, whole cell lysate 10 μg (Input).
    Lane 2: ab217673 IP in HeLa treated with 100 ng/ml nocodazole for 18 hours, whole cell lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab217673 in HeLa treated with 100 ng/ml nocodazole for 18 hours, whole cell lysate.

    Exposure time: 1 second.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

     

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab217673).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab217673).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab217673).

References

ab228854 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab228854.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up