Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-MTAP antibody [EPR6893] - BSA and Azide free (ab232417)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR6893] to MTAP - BSA and Azide free
  • Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-MTAP antibody [EPR6893] - BSA and Azide free
    See all MTAP primary antibodies

  • Description

    Rabbit monoclonal [EPR6893] to MTAP - BSA and Azide free

  • Host species

    Rabbit

  • Specificity

    The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: HeLa, 293T, HT29, C6, RAW 264.7, and NIH 3T3 cell lysates. ICC/IF: HeLa cells. Flow Cyt: HeLa cells. IHC-P: Human kidney, mouse kidney, and human lung carcinoma tissue.

  • General notes

    ab232417 is the carrier-free version of ab126770. 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

    ab232417 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Western blot - Anti-MTAP antibody [EPR6893] - BSA and Azide free (ab232417)
    Western blot - Anti-MTAP antibody [EPR6893] - BSA and Azide free (ab232417)
    All lanes : Anti-MTAP antibody [EPR6893] (ab126770) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : MTAP knockout HeLa cell lysate
    Lane 3 : HT-29 cell lysate
    Lane 4 : A549 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 31 kDa
    Observed band size: 32 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 (ab126770).

    anes 1- 4: Merged signal (red and green). Green - ab126770 observed at 32 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

    ab126770 Anti-MTAP antibody [EPR6893] was shown to specifically react with MTAP in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265272 (knockout cell lysate ab257194) was used. Wild-type and MTAP knockout samples were subjected to SDS-PAGE. ab126770 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 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 10000 dilution for 1 hour at room temperature before imaging.

     

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTAP antibody [EPR6893] - BSA and Azide free (ab232417)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTAP antibody [EPR6893] - BSA and Azide free (ab232417)

    Formalin-fixed, paraffin-embedded human kidney tissue stained for MTAP with unpurified ab126770 (1/50 dilution) in immunohistochemical analysis.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126770).

    Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

  • Immunoprecipitation - Anti-MTAP antibody [EPR6893] - BSA and Azide free (ab232417)
    Immunoprecipitation - Anti-MTAP antibody [EPR6893] - BSA and Azide free (ab232417)

    ab126770 (purified) at 1:50 dilution (2µg) immunoprecipitating MTAP in HT-29 whole cell lysate.
    Lane 1 (input): HT-29 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate 10ug
    Lane 2 (+): ab126770 & HT-29 whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab126770 in HT-29 whole cell lysate
    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126770).

  • Flow Cytometry - Anti-MTAP antibody [EPR6893] - BSA and Azide free (ab232417)
    Flow Cytometry - Anti-MTAP antibody [EPR6893] - BSA and Azide free (ab232417)
    Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling MTAP with purified ab126770 at 1:90 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126770).

  • Immunocytochemistry/ Immunofluorescence - Anti-MTAP antibody [EPR6893] - BSA and Azide free (ab232417)
    Immunocytochemistry/ Immunofluorescence - Anti-MTAP antibody [EPR6893] - BSA and Azide free (ab232417)

    Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labeling MTAP with Purified ab126770 at 1/250 dilution. Cells were fixed with 4% PFA and permeabilized with 0.1% tritonX-100. ab150077 Goat anti rabbit IgG (Alexa Fluor® 488) at 1/1000 was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126770).

  • Flow Cytometry - Anti-MTAP antibody [EPR6893] - BSA and Azide free (ab232417)
    Flow Cytometry - Anti-MTAP antibody [EPR6893] - BSA and Azide free (ab232417)

    Overlay histogram showing HeLa cells stained with unpurified ab126770 (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 (ab126770, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126770).

  • OI-RD Scanning - Anti-MTAP antibody [EPR6893] - BSA and Azide free (ab232417)
    OI-RD Scanning - Anti-MTAP antibody [EPR6893] - BSA and Azide free (ab232417)
    Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126770).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTAP antibody [EPR6893] - BSA and Azide free (ab232417)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MTAP antibody [EPR6893] - BSA and Azide free (ab232417)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human lung carcinoma tissue sections labeling MTAP with Purified ab126770 at 1:1000 dilution (0.89 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126770).

  • Anti-MTAP antibody [EPR6893] - BSA and Azide free (ab232417)
    Anti-MTAP antibody [EPR6893] - BSA and Azide free (ab232417)

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

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