Recombinant
RabMAb

Recombinant Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)

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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)
  • Suitable for: Dot blot, IHC-P, WB, ICC/IF, IP
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444]

  • Description

    Rabbit monoclonal [EPR18444] to ERK1 (phospho T202) + ERK2 (phospho T185)

  • Host species

    Rabbit

  • Specificity

    ab214036 does not react with a peptide containing ERK1 pY204 or ERK2 pY187

  • Tested applications

    Suitable for: Dot blot, IHC-P, WB, ICC/IF, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human ERK1 aa 150-250 (phospho T202 + Y204). The exact sequence is proprietary.
    Database link: P27361

  • 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

    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.

Properties

Applications

Our Abpromise guarantee covers the use of ab214036 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Dot blot Use at an assay dependent concentration.
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 44, 42 kDa (predicted molecular weight: 43, 41 kDa).
ICC/IF 1/100.
IP 1/40.

Images

  • Western blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)
    Western blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)
    All lanes : Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036) at 1/1000 dilution

    Lane 1 : Untreated Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
    Lane 2 : Jurkat (Human T cell leukemia cell line from peripheral blood) treated with 200 ng/ml PMA for 30 minutes whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 43, 41 kDa
    Observed band size: 42,44 kDa
    why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunocytochemistry/ Immunofluorescence - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)
    Immunocytochemistry/ Immunofluorescence - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)

    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).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)

    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.

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

  • Immunoprecipitation - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)
    Immunoprecipitation - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)

    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 1/10,000 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.

  • Dot Blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)
    Dot Blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)

    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.

  • Western blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)
    Western blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)
    All lanes : Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036) at 1/1000 dilution

    Lane 1 : Untreated NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
    Lane 2 : NIH/3T3 (Mouse embryonic fibroblast cell line) treated with 50 ng/ml PDGF for 40 minutes whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 43, 41 kDa
    Observed band size: 42,44 kDa why is the actual band size different from the predicted?


    Exposure time: 2 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)

    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.

    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] (ab214036)
    Immunocytochemistry/ Immunofluorescence - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)

    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.

  • Immunoprecipitation - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)
    Immunoprecipitation - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)

    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 1/10,000 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.

  • Western blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)
    Western blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)
    All lanes : Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036) at 1/1000 dilution

    Lane 1 : Untreated PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma cell line) treated with 200 ng/ml NGF for 4 days whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 43, 41 kDa
    Observed band size: 42,44 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)

    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.

    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] (ab214036)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)

    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.

    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] (ab214036)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)

    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.

    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] (ab214036)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)

    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.

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

References

Publishing research using ab214036? Please let us know so that we can cite the reference in this datasheet.

ab214036 has been referenced in 7 publications.

  • Liu D  et al. microRNA-155 Modulates Hepatic Stellate Cell Proliferation, Apoptosis, and Cell Cycle Progression in Rats With Alcoholic Hepatitis via the MAPK Signaling Pathway Through Targeting SOCS1. Front Pharmacol 11:270 (2020). PubMed: 32317960
  • Zhao J  et al. Long non-coding RNA ANRIL down-regulates microRNA-7 to protect human trabecular meshwork cells in an experimental model for glaucoma. Eur Rev Med Pharmacol Sci 23:3173-3182 (2019). PubMed: 31081068
  • Zhu AD  et al. lncRNA-ATB promotes viability, migration, and angiogenesis in human microvascular endothelial cells by sponging microRNA-195. J Cell Biochem N/A:N/A (2019). PubMed: 30983015
  • Gao S  et al. miR-9 depletion suppresses the proliferation of osteosarcoma cells by targeting p16. Int J Oncol 54:1921-1932 (2019). PubMed: 31081054
  • Zhou ZW  et al. Effect of willed movement training on neurorehabilitation after focal cerebral ischemia and on the neural plasticity-associated signaling pathway. Mol Med Rep 17:1173-1181 (2018). PubMed: 29115485
  • Xu J  et al. Combination of curcumin and vagus nerve stimulation attenuates cerebral ischemia/reperfusion injury-induced behavioral deficits. Biomed Pharmacother 103:614-620 (2018). WB ; Rat . PubMed: 29677548
  • Liu Z  et al. Astrocytes induce proliferation of oligodendrocyte progenitor cells via connexin 47-mediated activation of the ERK/Id4 pathway. Cell Cycle 16:714-722 (2017). PubMed: 28278052

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