Anti-Vimentin (phospho S72) antibody [EP1070Y] (ab52944)

Overview

  • Product name
    Anti-Vimentin (phospho S72) antibody [EP1070Y]
    See all Vimentin primary antibodies
  • Description
    Rabbit monoclonal [EP1070Y] to Vimentin (phospho S72)
  • Host species
    Rabbit
  • Specificity
    ab52944 only detects Vimentin phosphorylated at Serine 72.
  • Tested applications
    Suitable for: WB, IP, Dot blotmore details
    Unsuitable for: ICC/IF or IHC-P
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Vimentin aa 50-150 (phospho ). The exact sequence is proprietary.

  • Positive control
    • HeLa cell lysate (untreated and Calyculin A treated); NIH/3T3; C6 lysates.
  • General notes

    A trial size is available to purchase for this antibody.

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab52944 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 1/10000. Detects a band of approximately 54 kDa (predicted molecular weight: 54 kDa).
IP 1/20.

For unpurified use at 1:40. 

Dot blot Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for ICC/IF or IHC-P.
  • 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

    • ab52944 (purified) at 1:20 dilution (2µg) immunoprecipitating Vimentin in HeLa starved 24 hours, then treated with 100nM Calyculin A for 30 minutes whole cell lysate.
      Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) starved 24 hours,then treated with 100nM Calyculin A for 30 minutes whole cell lysate 10ug
      Lane 2 (+): ab52944 & HeLa starved 24 hours, then treated with 100nM Calyculin A for 30 minutes whole cell lysate
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab52944 in HeLa starved 24 hours,then treated with 100nM Calyculin A for 30 minutes whole cell lysate
      For western blotting, VeriBlot for IP secondary antibody (HRP) (ab131366) was used as the secondary antibody at 1:1000 dilution.
      Blocking and diluting buffer: 5% NFDM/TBST.
    • All lanes : Anti-Vimentin (phospho S72) antibody [EP1070Y] (ab52944) at 1/10000 dilution (purified)

      Lane 1 : Untreated with C6 (Rat glial tumor glial cell) whole cell lysates
      Lane 2 : C6 (Rat glial tumor glial cell) starved for 24 hours,then treated with 100ng Calyculin A for 60 minutes whole cell lysates

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 54 kDa



      Blocking and diluting buffer : 5% NFDM/TBST
    • All lanes : Anti-Vimentin (phospho S72) antibody [EP1070Y] (ab52944) at 1/10000 dilution (purified)

      Lane 1 : Untreated with HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
      Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) starved for 24 hours,then treated with 100nM Calyculin A for 30 minutes whole cell lysates

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 54 kDa



      Blocking and diluting buffer : 5% NFDM/TBST

    • All lanes : Anti-Vimentin (phospho S72) antibody [EP1070Y] (ab52944) at 1/10000 dilution (purified)

      Lane 1 : Untreated with NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates
      Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) treated with 100ng/ml Calyculin A for 30 minutes whole cell lysates

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 54 kDa



      Blocking and diluting buffer : 5% NFDM/TBST
    • Primary antibody dilution: 1/1000
      Secondary antibody: goat anti-rabbit IgG, (H+L), peroxidase conjugated (1/2500 dilution)
      Blocking & dilutiing buffer: 5% NFDM/TBST

      Lane 1 sample: Vimentin (pS72) phospho peptide
      Lane 2 sample: Vimentin non-phospho peptide

      Exposure time: 3 minutes

    • All lanes : Anti-Vimentin (phospho S72) antibody [EP1070Y] (ab52944) at 1/1000000 dilution

      Lane 1 : HeLa cell lysate untreated
      Lane 2 : HeLa cell lysate treated with Calyculin A

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : goat anti-rabbit HRP at 1/2000 dilution

      Predicted band size: 54 kDa
      Observed band size: 54 kDa

    References

    This product has been referenced in:
    • Goto H  et al. Phospho-Specific Antibody Probes of Intermediate Filament Proteins. Methods Enzymol 568:85-111 (2016). Read more (PubMed: 26795468) »
    • Mähönen AJ  et al. Culture medium induced vimentin reorganization associates with enhanced baculovirus-mediated gene delivery. J Biotechnol 145:111-9 (2010). Read more (PubMed: 19903502) »

    See all 2 Publications for this product

    Customer reviews and Q&As

    The lab has confirmed that the recommended dilution for WB is 1:1000000.

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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