Recombinant
RabMAb

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

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Overview

  • Product name

    Anti-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)

  • Host species

    Rabbit

  • Specificity

    This antibody detects Erk1 phosphorylated at Threonine 202 and Tyrosine 204 and Erk2 phosphorylated at Threonine 185 and Tyrosine 187.

  • Tested applications

    Suitable for: WB, IP, Dot blot, ELISAmore details
    Unsuitable for: Flow Cyt,ICC/IF or IHC-P

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human Erk1 (pT202/pY204) + Erk2 (pT185/pY187). The exact sequence is proprietary. 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 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 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.
Dot blot Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF or IHC-P.

    • Images

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

      Dot blot analysis of single phospho peptide pT202 (Lane 1), single phospho peptide pY204 (Lane 2), double 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.

    • Western blot - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299)
      Western blot - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299)
      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
      All lanes : 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.

    • Western blot - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299)
      Western blot - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299)
      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
      All lanes : 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.

    • Western blot - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299)
      Western blot - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299)
      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
      All lanes : 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.

    • Western blot - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299)
      Western blot - Anti-Erk1 (pT202/pY204) + Erk2 (pT185/pY187) antibody [EP197Y] (ab76299)
      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
      All lanes : 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.

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

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

      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.

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

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

      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

    Publishing research using ab76299? Please let us know so that we can cite the reference in this datasheet.

    ab76299 has been referenced in 41 publications.

    • Yuan C  et al. OAB-14, a bexarotene derivative, improves Alzheimer's disease-related pathologies and cognitive impairments by increasing ß-amyloid clearance in APP/PS1 mice. Biochim Biophys Acta Mol Basis Dis 1865:161-180 (2019). PubMed: 30389579
    • Sun C  et al. Astrocytic miR-324-5p is essential for synaptic formation by suppressing the secretion of CCL5 from astrocytes. Cell Death Dis 10:141 (2019). PubMed: 30760705
    • Wen Q  et al. FoxO3a inhibiting expression of EPS8 to prevent progression of NSCLC: A new negative loop of EGFR signaling. EBioMedicine 40:198-209 (2019). PubMed: 30738830
    • Li X  et al. The Specific Inhibition of SOD1 Selectively Promotes Apoptosis of Cancer Cells via Regulation of the ROS Signaling Network. Oxid Med Cell Longev 2019:9706792 (2019). PubMed: 30911355
    • Xiang XJ  et al. Mitochondrial methionine sulfoxide reductase B2 links oxidative stress to Alzheimer's disease-like pathology. Exp Neurol 318:145-156 (2019). PubMed: 31078523
    View all Publications for this product

    Customer reviews and Q&As

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    Question

    Thank you for your e-mail informing me about the change in the
    specificity of ab76299.
    I wish avail of your offer to receive a credit note. The original order number is xxx. I would greatly appreciate if the order number could be addressed to me at the address below. If you have any queries please do not hesitate to contact me either at
    this e-mail address or using the following phone number: xxx

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    Answer

    Thank you for contacting us.

    Your credit note ID is xxx.

    I am sorry that this antibody had a mistake on its datasheet - which we have corrected in the meantime.I have asked our Finance department to issue a credit note for you as you had requested. Also, I have added your contact information per your instructions.The credit note may be used in one of the following ways:

    (1) Redeemed against the original invoice if this hasn't already been paid.
    (2) Held on the account for use against a future order.
    (3) A full refund can be offered where no other invoices are outstanding.

    Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.

    To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website.

    The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.

    I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice.

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    Question

    Thank you for sharing this very important piece of
    information with us.
    There is disappointment in the the lab regarding this error. In
    any case,
    the original order number is xxx. Please send me a receipt
    or an invoice indicating the refund.

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    Answer

    Thank you for contacting us.

    Your credit note ID for the refund is xxx.

    We are very sorry aboutthe mistakemade for the target name of the antibody. This has now been corrected.

    If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.

    Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.

    The credit note ID is for your reference only and does not automatically guarantee the credit.

    We are very sorry for the oversight andhope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

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