Anti-Acidic Cytokeratin antibody [AE1] (ab3117)

Overview

  • Product nameAnti-Acidic Cytokeratin antibody [AE1]See all Acidic Cytokeratin primary antibodies ...
  • Description
    Mouse monoclonal [AE1] to Acidic Cytokeratin
  • SpecificityThis antibody reacts with MCF-7, RT 112 and A 431 cultured cell lines.
  • Tested applicationsICC, IHC-P, IHC-Fr, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Human, Monkey
    Predicted to work with: Mouse, Rat, Rabbit, Chicken, Cow
  • Immunogen

    Human epidermal keratin.

  • Positive control
    • Skin. Squamous cellcarcinoma.
  • General notes


    Twenty human keratins are resolved with two-dimensional gel electrophoresis into acidic (pI <5.7) and basic (pI >6.0) subfamilies. The acidic keratins have molecular weights of 56.5, 55, 51, 50, 50', 48, 46, 45, and 40kDa. The basic keratins have molecular weights of 65 - 67, 64, 59, 58, 56 and 52kDa. Members of acidic and basic subfamilies are found together in pairs. The composition of keratin pairs varies with cell type, differentiation status and environment. Many studies have shown the usefulness of keratins as markers in cancer research and tumor diagnosis.

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer10mM PBS, pH7.4, 0.2%BSA, 0.09% sodium azide
  • Concentration information loading...
  • PurityProtein G purified
  • Primary antibody notes Twenty human keratins are resolved with two-dimensional gel electrophoresis into acidic (pI <5.7) and basic (pI >6.0) subfamilies. The acidic keratins have molecular weights of 56.5, 55, 51, 50, 50', 48, 46, 45, and 40kDa. The basic keratins have molecular weights of 65 - 67, 64, 59, 58, 56 and 52kDa. Members of acidic and basic subfamilies are found together in pairs. The composition of keratin pairs varies with cell type, differentiation status and environment. Many studies have shown the usefulness of keratins as markers in cancer research and tumor diagnosis.
  • Clonality Monoclonal
  • Clone numberAE1
  • IsotypeIgG1
  • Research Areas

Applications

Our Abpromise guarantee covers the use of ab3117 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
ICC Use at an assay dependent dilution.
IHC-P Use at an assay dependent dilution. For staining of formalin-fixed tissues, digest sections with trypsin at 1mg/ml PBS, 10 min at 37°C or Protease XXV at 1mg/ml PBS for 5minutes at 37°C. This antibody is an excellent marker for distinguishing carcinomas from non-epithelial tumours. This antibody used alongside ab9287 has also been used extensively for distinguishing carcinoma from non-carcinoma.
IHC-Fr Use at an assay dependent dilution.
Flow Cyt Use 1µg for 106 cells.
ICC/IF Use a concentration of 5 µg/ml.
  • Application notesIs unsuitable for WB.
  • Target

    Anti-Acidic Cytokeratin antibody [AE1] images

    • ab3117 (1/100) staining formalin-fixed, paraffin embedded human colon tissue sections. Antigen retrieval performed by digestion with trypsin at 1mg/ml for 10min at 37oC.
    • ICC/IF image of ab3117 stained MCF7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab3117, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    • Overlay histogram showing A431 cells stained with ab3117 (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. The cells were then incubated with the antibody (ab3117, 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 IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    References for Anti-Acidic Cytokeratin antibody [AE1] (ab3117)

    This product has been referenced in:
    • Chai J  et al. CCN1 induces a reversible epithelial-mesenchymal transition in gastric epithelial cells. Lab Invest 90:1140-51 (2010). ICC/IF . Read more (PubMed: 20458273) »

    See 1 Publication for this product

    Product Wall

    Leider hat es sich herausgestellt, dass die Information doch nicht so leicht zu beschaffen war und dann leider noch nicht mal besonders reichlich ist: Dieser Antikörper wird mit den meisten sauren Type I Keratine reagieren. Leider konnte man mir nicht ...

    Read More
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunocytochemistry/ Immunofluorescence
    Sample Human Cell (gastric adenocarcinoma)
    Specification gastric adenocarcinoma
    Fixative Methanol
    Permeabilization Yes - 0.1% Triton
    Blocking step DAKO serum free protein blocker as blocking agent for 20 minute(s) · Concentration: 0.25% · Temperature: 28°C
    Username

    Dr. J Chai

    Verified customer

    Submitted Apr 13 2012

    Thank you for your phone call. I regret to inform you that the antibody's originator has informed me that ab3117 does not work in Western blotting. There was an error on the online datasheet which I have since updated. So, I unfortunately do not know w...

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