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RabMAb

Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299)

Overview

  • Product nameAnti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y]
    See all Erk1 (pT202/pY204) + Erk2 (pT185/pY187) primary antibodies
  • Description
    Rabbit monoclonal [EP197Y] to Erk1 (pT202/pY204) + Erk2 (pT185/pY187)
  • SpecificityThis antibody detects Erk1 phosphorylated at Threonine 202 and Tyrosine 204 and Erk2 phosphorylated at Threonine 185 and Tyrosine 187.
  • Tested applicationsWB, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Erk1 (pT202/pY204) + Erk2 (pT185/pY187). Swissprot Numbers: P27361, P28482.

  • Positive control
    • WB: A431 (treated with EGF), SH-SY5Y (treated with NGF), PC-12 (treated with NGF), HeLa and NIH/3T3 whole cell lysates. IP: A431 (treated with EGF) and PC-12 (treated with NGF) whole cell lysates.
  • General notesProduced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

Properties

Applications

Our Abpromise guarantee covers the use of ab76299 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
WB 1/5000 - 1/10000. Detects a band of approximately 43, 42 kDa (predicted molecular weight: 43, 41 kDa).
IP 1/80.
  • Application notesIs unsuitable for Flow Cyt,ICC/IF or IHC-P.
  • Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] images

    • Dot blot analysis of single phospho peptide pT202 (Lane 1), single phospho peptide pY204 (Lane 2), souble phospho peptide pT202/pY204 (Lane 3) and non-phospho peptide (Lane 4) labelling Erk1 (pT202/pY204) + Erk2 (p185/pY187) with ab76299 at a dilution of 1/1000. A peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody at a dilution of 1/1000.

      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure time: 3 minutes.

    • All lanes : Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299) at 1/5000 dilution

      Lane 1 : A431 whole cell lysate - untreated
      Lane 2 : A431 whole cell lysate - treated with 100ng/ml EGF for 10 minutes

      Lysates/proteins at 10 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

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


      Exposure time : 3 minutes

      Blocking and dilution buffer: 5% NFDM/TBST.

    • All lanes : Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299) at 1/10000 dilution

      Lane 1 : SH-SY5Y whole cell lysate - untreated
      Lane 2 : SH-SY5Y whole cell lysate - treated with 100ng/ml NGF for 10 minutes

      Lysates/proteins at 10 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 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 : 15 seconds

      Blocking and dilution buffer: 5% NFDM/TBST.

    • All lanes : Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299) at 1/10000 dilution

      Lane 1 : PC-12 whole cell lysate - untreated
      Lane 2 : PC-12 whole cell lysate - treated with 100ng/ml NGF for 10 minutes

      Lysates/proteins at 10 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 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 : 15 seconds

      Blocking and dilution buffer: 5% NFDM/TBST.

    • All lanes : Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299) at 1/10000 dilution

      Lane 1 : HeLa whole cell lysate - untreated
      Lane 2 : HeLa whole cell lysate - treated with Lambda Phosphatase

      Lysates/proteins at 10 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 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 and dilution buffer: 5% NFDM/TBST.

    • Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299) at 1/10000 dilution + NIH/3T3 whole cell lysate at 10 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 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 and dilution buffer: 5% NFDM/TBST.

    • ab76299 at 1/80 immunoprecipitating Erk1 (pT202/pY204) + Erk2 (pT185/pY187) in A431 whole cell lysate.

      Lane 1 (input): A431 whole cell lysate - treated with 100ng/ml EGF for 10 min (10µg).

      Lane 2 (+): ab76299 + A431 whole cell lysate - treated with 100ng/ml EGF for 10 min.

      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab76299 in A431 whole cell lysate - treated with 100ng/ml EGF for 10 min.

      For western blotting, ab76299 was used at a dilution of 1/1000 and a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG, was used as the secondary antibody (1/1500).

      Blocking and dilution buffer: 5% NFDM/TBST.

    • ab76299 at 1/80 immunoprecipitating Erk1 (pT202/pY204) + Erk2 (pT185/pY187) in PC-12 whole cell lysate.

      Lane 1 (input): PC-12 whole cell lysate - treated with 100ng/ml NGF for 10 min (10µg).

      Lane 2 (+): ab76299 + PC-12 whole cell lysate - treated with 100ng/ml NGF for 10 min.

      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab76299 in PC-12 whole cell lysate - treated with 100ng/ml NGF for 10 min.

      For western blotting, ab76299 was used at a dilution of 1/1000 and a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG, was used as the secondary antibody (1/1500).

      Blocking and dilution buffer: 5% NFDM/TBST.

    • ELISA analysis of various phospho and non-phospho peptides (0-250 ng/ml) labelling Erk1 (pT202/pY204) + Erk2 (pT185/pY187) with ab76299 at a dilution of 1/1000. An Alkaline Phosphatase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/2500).

      ab76299 has stronger affinity for the double phospho peptide Y204/Y187 than to single phospho peptides T202 or Y204.

    References for Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299)

    This product has been referenced in:
    • Jin H  et al. Matrine activates PTEN to induce growth inhibition and apoptosis in V600EBRAF harboring melanoma cells. Int J Mol Sci 14:16040-57 (2013). WB ; Human . Read more (PubMed: 23912239) »
    • Wang Y  et al. Hypoxia-Inducible Factor-1alpha and MAPK Co-Regulate Activation of Hepatic Stellate Cells upon Hypoxia Stimulation. PLoS One 8:e74051 (2013). WB ; Human . Read more (PubMed: 24040163) »

    See all 6 Publications for this product

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