Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-MLK3 antibody [EP1460Y] - BSA and Azide free (ab239840)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP1460Y] to MLK3 - BSA and Azide free
  • Suitable for: ICC, WB
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-MLK3 antibody [EP1460Y] - BSA and Azide free
    See all MLK3 primary antibodies

  • Description

    Rabbit monoclonal [EP1460Y] to MLK3 - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ICC
    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: A549, HAP1, A431, HepG2, and HeLa cell lysates; ICC: HeLa cells.

  • General notes

    ab239840 is the carrier-free version of ab51068 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

    Ab239840 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Mouse and Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    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.

    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

Images

  • Western blot - Anti-MLK3 antibody [EP1460Y] - BSA and Azide free (ab239840)
    Western blot - Anti-MLK3 antibody [EP1460Y] - BSA and Azide free (ab239840)
    All lanes : Anti-MLK3 antibody [EP1460Y] (ab51068) at 1/1000 dilution (Purified)

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 2 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate
    Lane 3 : A431 (Human epidermoid carcinoma epithelial cell) whole cell lysate

    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: 93 kDa



    This data was developed using ab51068, the same antibody clone in a different buffer formulation.

    Blocking buffer: 5% NFDM/TBST

  • Immunocytochemistry - Anti-MLK3 antibody [EP1460Y] - BSA and Azide free (ab239840)
    Immunocytochemistry - Anti-MLK3 antibody [EP1460Y] - BSA and Azide free (ab239840)

    This data was developed using ab51068 the same antibody clone in a different buffer formulation.
    Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling MLK3 with Purified ab51068 at 1:50 dilution (3.52 µg/ml). Cells were fixed in 100% Methanol and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor® 488,ab150077) was used as the secondary antibody at 1:1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Western blot - Anti-MLK3 antibody [EP1460Y] - BSA and Azide free (ab239840)
    Western blot - Anti-MLK3 antibody [EP1460Y] - BSA and Azide free (ab239840)
    All lanes : Anti-MLK3 antibody [EP1460Y] (ab51068) at 1/5000 dilution

    Lane 1 : Wild-type A549 cell lysate
    Lane 2 : MAP3K11 knockout A549 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 93 kDa
    Observed band size: 105 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 (ab51068).

    Lanes 1- 2: Merged signal (red and green). Green - ab51068 observed at 105 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab51068 was shown to react with MLK3 in wild-type A549 cells in western blot. Loss of signal was observed when knockout cell line ab267169 (knockout cell lysate ab257519) was used. Wild-type A549 and MAP3K11 knockout A549 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. ab51068 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 5000 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.

  • Western blot - Anti-MLK3 antibody [EP1460Y] - BSA and Azide free (ab239840)
    Western blot - Anti-MLK3 antibody [EP1460Y] - BSA and Azide free (ab239840)
    All lanes : Anti-MLK3 antibody [EP1460Y] (ab51068) at 1/5000 dilution

    Lane 1 : Wild-type A549 cell lysate
    Lane 2 : MAP3K11 knockout A549 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 93 kDa
    Observed band size: 105 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 (ab51068).

    Lanes 1- 2: Merged signal (red and green). Green - ab51068 observed at 105 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab51068 was shown to react with MLK3 in wild-type A549 cells in western blot. Loss of signal was observed when knockout cell line ab267168 (knockout cell lysate ab257518) was used. Wild-type A549 and MAP3K11 knockout A549 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. ab51068 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 5000 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.

  • Western blot - Anti-MLK3 antibody [EP1460Y] - BSA and Azide free (ab239840)
    Western blot - Anti-MLK3 antibody [EP1460Y] - BSA and Azide free (ab239840)

    This data was developed using the same antibody clone in a different buffer formulation (ab51068).

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: MLK3 knockout HAP1 cell lysate (20 µg) 
    Lane 3: A431 cell lysate (20 µg)
    Lane 4: HeLa cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab51068 observed at 105 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab51068 was shown to specifically react with MLK3 when MLK3 knockout samples were used. Wild-type and MLK3 knockout samples were subjected to SDS-PAGE. ab51068 and ab8245 (loading control to GAPDH) were diluted 1/5000 and 1/1000 respectively and incubated overnight at 4°C. 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 1/10000 dilution for 1 hour at room temperature before imaging.

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