Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-MUC1 antibody [EP1024Y] - Low endotoxin, Azide free (ab218998)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1024Y] to MUC1 - Low endotoxin, Azide free
  • Suitable for: Flow Cyt, IP, ICC/IF, WB
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-MUC1 antibody [EP1024Y] - Low endotoxin, Azide free
    See all MUC1 primary antibodies

  • Description

    Rabbit monoclonal [EP1024Y] to MUC1 - Low endotoxin, Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt, IP, ICC/IF, WBmore details

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa, T47D, MCF7 and A549 cell lysates; Human kidney, human breast carcinoma, human thyroid carcinoma and human colon cancer lysates; Human fetal lung lysate; Rat liver lysate and mouse liver lysate. ICC/IF: MCF7 cells. Flow Cyt: T47D and A549 cells.

  • General notes

    ab218998 is a carrier-free antibody designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

    Isoform 7 of MUC1 behaves as a receptor and binds the secreted isoform 5. The binding induces the phosphorylation of the isoform 7, alters cellular morphology and initiates cell signaling. The mouse and rat recommendation is based on WB results.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

Properties

Images

  • Western blot - Anti-MUC1 antibody [EP1024Y] - Low endotoxin, Azide free (ab218998)
    Western blot - Anti-MUC1 antibody [EP1024Y] - Low endotoxin, Azide free (ab218998)
    All lanes : Anti-MUC1 antibody [EP1024Y] (ab45167) at 1/1000 dilution

    Lane 1 : MCF7 (Human breast adenocarcinoma cell line) cell lysate
    Lane 2 : T-47D (Human ductal breast epithelial tumor cell line) cell lysate
    Lane 3 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
    Lane 4 : MUC1 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Observed band size: 24 kDa
    why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab45167).

    Lanes 1- 4: Merged signal (red and green). Green - ab45167 observed at 24 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab45167 was shown to react with MUC1 in Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) cells in western blot. Loss of signal was observed when knockout cell line ab255412 (knockout cell lysate ab263764) was used. Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) and MUC1 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab45167 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-MUC1 antibody [EP1024Y] - Low endotoxin, Azide free (ab218998)
    Immunocytochemistry/ Immunofluorescence - Anti-MUC1 antibody [EP1024Y] - Low endotoxin, Azide free (ab218998)

    Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma cell line) cells labeling MUC1 with purified ab45167 at 1/500 dilution (0.2μg/ml). Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, a goat anti-rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1/1000 dilution. Ab195889, an anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used as counterstain at 1/200 dilution (2.5 μg/ml). The negative control is PBS instead of the primary antibody. Nuclei counterstained with DAPI (blue). 

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45167).

  • Immunoprecipitation - Anti-MUC1 antibody [EP1024Y] - Low endotoxin, Azide free (ab218998)
    Immunoprecipitation - Anti-MUC1 antibody [EP1024Y] - Low endotoxin, Azide free (ab218998)

    ab45167 (purified) at 1/20 dilution (2ug) immunoprecipitating MUC1 in Human fetal lung lysate.

    Lane 1 (input): Human fetal lung lysate 10ug

    Lane 2 (+): ab45167 + Human fetal lung lysate 10ug

    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab45167 in Human fetal lung lysate

    For western blotting, ab131366 VeriBlot for IP (HRP) was used for detection at 1/1000 dilution.

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting 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 (ab45167).

  • Flow Cytometry - Anti-MUC1 antibody [EP1024Y] - Low endotoxin, Azide free (ab218998)
    Flow Cytometry - Anti-MUC1 antibody [EP1024Y] - Low endotoxin, Azide free (ab218998)

    Flow Cytometry analysis of A549 (human lung carcinoma cell line) cells labeling MUC1 with purified ab45167 at 1/20 dilution (10 ug/ml). Cells were fixed with 4% paraformaldehyde. A goat anti-rabbit IgG (Alexa Fluor® 488) was used as the secondary antibody at 1/2000 dilution. Black - Isotype control, Rabbit monoclonal IgG. Blue - unlabeled control, cells without incubation with primary antibody and secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45167).

  • Flow Cytometry - Anti-MUC1 antibody [EP1024Y] - Low endotoxin, Azide free (ab218998)
    Flow Cytometry - Anti-MUC1 antibody [EP1024Y] - Low endotoxin, Azide free (ab218998)

    Flow cytometry analysis of T47D (human mammary gland ductal carcinoma) cells labelling MUC1 with unpurified ab45167 (pink) at a dilution of 1/150. Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG was used as the secondary antibody. Rabbit monoclonal IgG (ab172730) was used as the isotype control (green).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab45167).

  • Anti-MUC1 antibody [EP1024Y] - Low endotoxin, Azide free (ab218998)
    Anti-MUC1 antibody [EP1024Y] - Low endotoxin, Azide free (ab218998)

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