Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR17526] to ERK1 + ERK2 - BSA and Azide free
  • Suitable for: ICC, Flow Cyt, IP, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free
    See all ERK1 + ERK2 primary antibodies

  • Description

    Rabbit monoclonal [EPR17526] to ERK1 + ERK2 - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    IP
    Rat
    WB
    Recombinant fragment
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: HeLa cell lysate. ICC/IF: HeLa cells. Flow Cyt: A431 cells. IP. PC-12 whole cell extract.

  • General notes

    Ab218017 is the carrier-free version of ab184699. 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

    ab218017 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-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)
    Western blot - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)
    All lanes : Anti-ERK1 + ERK2 antibody [EPR17526] (ab184699) at 1/10000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : MAPK1 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 43, 41 kDa
    Observed band size: 44 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 (ab184699).

    Lanes 1-2: Merged signal (red and green). Green - ab184699 observed at 44 kDa. Red - loading control ab8245 observed at 37 kDa.

     ab184699 Anti-ERK1 + ERK2 antibody [EPR17526] was shown to specifically react with ERK2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265052 (knockout cell lysate ab257525) was used. Wild-type and ERK2 knockout samples were subjected to SDS-PAGE. ab184699 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 10000 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 20000 dilution for 1 hour at room temperature before imaging.

     

     

  • Immunocytochemistry - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)
    Immunocytochemistry - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling ERK1 + ERK2 with ab184699 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing both nuclear and cytoplasmic staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows:
    -ve control 1: ab184699 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184699).

  • Immunoprecipitation - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)
    Immunoprecipitation - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)

    ERK1 + ERK2 were immunoprecipitated from 1mg of PC-12 (Rat adrenal gland pheochromocytoma) whole cell extract with ab184699 at 1/70 dilution. Western blot was performed from the immunoprecipitate using ab184699 at 1/5000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

    Lane 1: PC-12 whole cell extract. Lane 2: PBS instead of PC-12 whole cell extract.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    44kDa band represents ERK1. 42kDa band represents ERK2.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184699).

  • Flow Cytometry - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)
    Flow Cytometry - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)

    Clone EPR17526 (ab218017) has been successfully conjugated by Abcam. This image was generated using Anti-ERK1 + ERK2 antibody [EPR17526] (PE). Please refer to ab212153 for protocol details.

    Overlay histogram showing A431 cells stained with ab212153 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 90% methanol at -20°C for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab212153, 1/2500 dilution) for 30 min at 22°C.

    Isotype control antibody (black line) was rabbit IgG (monoclonal) Phycoerythrin (ab209478) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 50 mW Yellow/Green laser (561nm) and 586/15 bandpass filter.

  • Immunocytochemistry - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)
    Immunocytochemistry - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017) This image is courtesy of an Abreview submitted by Kirk Mcmanus.

    Ab184699 staining ERK1 + ERK2 in HeLa cells by ICC/IF (Immunocytochemistry/Immunofluorescence). Cells were fixed with paraformaldehyde and permeabilized with 0.5% Triton X-100. Samples were incubated with primary antibody (1/500 in PBS) for 1 hour at 22°C. A Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184699).

  • Immunocytochemistry - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)
    Immunocytochemistry - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)

    Clone EPR17526 (ab218017) has been successfully conjugated by Abcam. This image was generated using Anti-ERK1 + ERK2 antibody [EPR17526] (Alexa Fluor® 647). Please refer to ab208881 for protocol details.

    ab208881 staining ERK1 + ERK2 in A431 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab208881 at a 1/100 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at a 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

     

  • Immunocytochemistry - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)
    Immunocytochemistry - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)

    Clone EPR17526 (ab218017) has been successfully conjugated by Abcam. This image was generated using Anti-ERK1 + ERK2 antibody [EPR17526] (Alexa Fluor® 488). Please refer to ab208564 for protocol details.

    ab208564 staining ERK1 + ERK2 in A431 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab208564 at a 1/100 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at a 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    This product also gave a positive signal under the same testing conditions in A431 cells fixed with 4% formaldehyde (10 min).

     

  • Flow Cytometry - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)
    Flow Cytometry - Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)

    Flow cytometric analysis of A431 (Human epidermoid carcinoma) cells labeling ERK1 + ERK2 with ab184699 at 1/440 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab184699).

  • Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)
    Anti-ERK1 + ERK2 antibody [EPR17526] - BSA and Azide free (ab218017)

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