Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18444] to ERK1 (phospho T202) + ERK2 (phospho T185) - BSA and Azide free
  • Suitable for: IHC-P, ICC/IF, IP, WB, Dot blot
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free

  • Description

    Rabbit monoclonal [EPR18444] to ERK1 (phospho T202) + ERK2 (phospho T185) - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, ICC/IF, IP, WB, Dot blotmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

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

  • Positive control

    • WB: Jurkat treated with 200 ng/ml PMA for 30 minutes whole cell lysate; NIH/3T3 treated with 50 ng/ml PDGF for 40 minutes whole cell lysate; PC-12 treated with 200 ng/ml NGF for 4 days whole cell lysate. IHC-P: Human breast, placenta, breast cancer and glioma tissues; mouse kidney tissue; rat spleen tissue. ICC/IF: Jurkat cells treated with PMA treatment (200 ng/ml, 30min). IP: Jurkat treated with 200 ng/ml PMA for 30 minutes cell lysate; PC-12 treated with 200 ng/ml NGF for 4 days cell lysate.

  • General notes

    Ab222493 is the carrier-free version of ab214036. 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

    ab222493 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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

  • Dot Blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)
    Dot Blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)

    Dot blot analysis of ERK1 (pT202) peptide (Lane 1), ERK1 (pT204) peptide (Lane 2), ERK1 (pT202 + pT204) peptide (Lane 3) and ERK1 non-phospho peptide (Lane 4) labelling ERK1 (pT202) with ab214036.

    Exposure time: 3 minutes.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214036).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)

    Immunohistochemical analysis of paraffin-embedded human breast tissue labeling ERK1 (phospho T202) + ERK2 (phospho T185) with ab214036 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nuclear staining on human normal breast tissue is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) 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 (ab214036).

    Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)

    Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling ERK1 (phospho T202) + ERK2 (phospho T185) with ab214036 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nuclear and weak cytoplasmic staining on scattered cells of human placenta is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) 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 (ab214036).

    Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)

    Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling ERK1 (phospho T202) + ERK2 (phospho T185) with ab214036 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nuclear and weak cytoplasmic staining on human breast tissue cancer is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) 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 (ab214036).

    Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)

    Immunohistochemical analysis of paraffin-embedded human glioma tissue labeling ERK1 (phospho T202) + ERK2 (phospho T185) with ab214036 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. 

    Nuclear and weak cytoplasmic staining on human glioma is observed [PMID:17487353].

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) 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 (ab214036).

    Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)

    Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling ERK1 (phospho T202) + ERK2 (phospho T185) with ab214036 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nuclear and weak cytoplasmic staining on scattered cells of mouse kidney is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) 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 (ab214036).

    Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)

    Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling ERK1 (phospho T202) + ERK2 (phospho T185) with ab214036 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Nuclear and weak cytoplasmic staining on scattered cells of rat spleen is observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) 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 (ab214036).

    Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)
    Immunocytochemistry/ Immunofluorescence - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cell line from peripheral blood) cells labeling -ERK1 (phospho T202) + ERK2 (phospho T185) with ab214036 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining increased after PMA treatment (200 ng/ml, 30min), and LP treatment decreased the PMA induced staining.

    The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

    The negative controls are as follows:-

    -ve control 1: ab214036 at 1/100 dilution followed by Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.

    -ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214036).

  • Immunocytochemistry/ Immunofluorescence - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)
    Immunocytochemistry/ Immunofluorescence - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cell line from peripheral blood) cells labeling ERK1 (phospho T202) + ERK2 (phospho T185) with ab214036 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining increased after PMA treatment (200 ng/ml, 30min), and LP treatment decreased the PMA induced staining. For the “pan” antibody, the signal is unchanged after PMA treatment (200 ng/ml, 30min), and LP treatment. The nuclear counterstain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214036).

  • Immunoprecipitation - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)
    Immunoprecipitation - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)

    ERK1 (phospho T202) + ERK2 (phospho T185) was immunoprecipitated from 0.35 mg of Jurkat (Human T cell leukemia cell line from peripheral blood) treated with 200 ng/ml PMA for 30 minutes whole cell lysate with ab214036 at 1/40 dilution.

    Western blot was performed from the immunoprecipitate using ab214036 at 1/1000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: Jurkat treated with 200 ng/ml PMA for 30 minutes whole cell lysate 10µg (Input).

    Lane 2: ab214036 IP in Jurkat treated with 200 ng/ml PMA for 30 minutes whole cell lysate.

    Lane 3: Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab214036 in Jurkat treated with 200 ng/ml PMA for 30 minutes whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 minutes.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214036).

  • Immunoprecipitation - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)
    Immunoprecipitation - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)

    ERK1 (phospho T202) + ERK2 (phospho T185) was immunoprecipitated from 0.35 mg of PC-12 (Rat adrenal gland pheochromocytoma cell line) treated with 200 ng/ml NGF for 4 days whole cell lysate with ab214036 at 1/40 dilution.

    Western blot was performed from the immunoprecipitate using ab214036 at 1/1000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

    Lane 1: PC-12 treated with 200 ng/ml NGF for 4 days whole cell lysate 10µg (Input).

    Lane 2: ab214036 IP in PC-12 treated with 200 ng/ml NGF for 4 days whole cell lysate.

    Lane 3: Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab214036 in PC-12 treated with 200 ng/ml NGF for 4 days whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 30 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214036).

  • Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)
    Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] - BSA and Azide free (ab222493)

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