Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-ERK1 antibody [EP4967] - BSA and Azide free (ab214169)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP4967] to ERK1 - BSA and Azide free
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-ERK1 antibody [EP4967] - BSA and Azide free
    See all ERK1 primary antibodies

  • Description

    Rabbit monoclonal [EP4967] to ERK1 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

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

  • Positive control

    • WB: HEK-293T, Jurkat, A375, A431 and HUVEC cell lysates. IHC-P: Human colon adenocarcinoma tissue. Flow Cyt: Jurkat cells.

  • General notes

    Ab214169 is the carrier-free version of ab109282. 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

    ab214169 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.

    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.

    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-ERK1 antibody [EP4967] - BSA and Azide free (ab214169)
    Western blot - Anti-ERK1 antibody [EP4967] - BSA and Azide free (ab214169)
    All lanes : Anti-ERK1 antibody [EP4967] (ab109282) at 1/1000 dilution

    Lane 1 : Wild-type HEK-293T cell lysate
    Lane 2 : MAPK3 knockout HEK-293T cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 44 kDa
    Observed band size: 43 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 (ab109282).

    Lanes 1- 2: Merged signal (red and green). Green - ab109282 observed at 43 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab109282 was shown to react with ERK1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266519 (knockout cell lysate ab257099) was used. Wild-type HEK-293T and MAPK3 knockout HEK-293T 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. ab109282 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.

  • Flow Cytometry - Anti-ERK1 antibody [EP4967] - BSA and Azide free (ab214169)
    Flow Cytometry - Anti-ERK1 antibody [EP4967] - BSA and Azide free (ab214169)

    Flow Cytometry analysis of Jurkat (human acute T cell leukemia) cells labeling ERK1 with purified ab109282 at 1/30 dilution (red). The secondary antibody was Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution. A Rabbit monoclonal IgG (Black) was used as the isotype control and cells without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109282).

  • Immunocytochemistry/ Immunofluorescence - Anti-ERK1 antibody [EP4967] - BSA and Azide free (ab214169)
    Immunocytochemistry/ Immunofluorescence - Anti-ERK1 antibody [EP4967] - BSA and Azide free (ab214169)

    Immunocytochemistry/Immunofluorescence analysis of Jurkat (human acute T cell leukemia) cells labelling ERK1 with purified ab109282 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

    Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109282).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 antibody [EP4967] - BSA and Azide free (ab214169)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 antibody [EP4967] - BSA and Azide free (ab214169)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colonic adenocarcinoma tissue labelling ERK1 with unpurified ab109282 at 1/100 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109282).

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Western blot - Anti-ERK1 antibody [EP4967] - BSA and Azide free (ab214169)
    Western blot - Anti-ERK1 antibody [EP4967] - BSA and Azide free (ab214169)

    This WB data was generated using the same anti-EKR1 antibody clone, EP4967, in a different buffer formulation (cat# ab109282).

    Lanes 1, 3 and 5: Wild-type HAP1 cell lysate (20 µg)
    Lanes 2, 4 and 6: ERK1 knockout HAP1 cell lysate (20 µg)
    Lanes 1 and 2: Green signal from target - ab109282 observed at 42 kDa 
    Lanes 3 and 4: Red signal from loading control - ab8226 observed at 42 kDa
    Lanes 5 and 6: Merged (red and green) signal
    ab109282 was shown to specifically react with ERK1 when ERK1 knockout samples were used.
    Wild-type and ERK1 knockout samples were subjected to SDS-PAGE. ab109282 and ab8226 (loading control to beta actin) were both diluted 1/1000 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 h at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 antibody [EP4967] - BSA and Azide free (ab214169)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 antibody [EP4967] - BSA and Azide free (ab214169)

    This IHC data was generated using the same anti-EKR1 antibody clone, EP4967, in a different buffer formulation (cat# ab109282).

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human melanoma tissue labelling ERK1 with purified ab109282 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Anti-ERK1 antibody [EP4967] - BSA and Azide free (ab214169)
    Anti-ERK1 antibody [EP4967] - BSA and Azide free (ab214169)

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