Recombinant
RabMAb

Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (ab201015)

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Overview

  • Product name
    Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401]
  • Description
    Rabbit monoclonal [EPR19401] to ERK1 (phospho T202) + ERK2 (phospho T185)
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, IP, IHC-P, WB, Dot blotmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human ERK2 aa 150-250 (phospho T185). The exact sequence is proprietary.
    Database link: P28482

  • Positive control
    • WB: Jurkat, treated with 200 ng/ml PMA for 30 minutes whole cell lysate; NIH/3T3, untreated and treated with 50 ng/ml PDGF for 40 minutes whole cell lysates; PC-12, untreated and treated with 200 ng/ml NGF for 4 days whole cell lysates. IHC-P: Human breast and glioma tissues. IP: NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate; PC-12 treated with 100ng/ml NGF for 10min whole cell lysate. ICC/IF:Jurkat
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab201015 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
ICC/IF Use at an assay dependent concentration.
IP 1/30.
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

IHC is recommended for human only.
WB 1/1000. Detects a band of approximately 44, 42 kDa (predicted molecular weight: 41 kDa).
Dot blot 1/1000.

Images

  • Western blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (ab201015)
    Western blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (ab201015)
    All lanes : Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (ab201015) 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 20 µg per lane.

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

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


    Exposure time: 15 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunocytochemistry/ Immunofluorescence - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (ab201015)
    Immunocytochemistry/ Immunofluorescence - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (ab201015)

    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) and ERK2 (phospho T185)

    ERK1 (phospho T202) + ERK2 (phospho T185) with ab201015 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing staining on M phase cells (PMID:26529125). After PMA treatment (200 ng/ml, 30min), the staining was increased, and LP treatment decreased the PMA induced staining.

    The nuclear counter stain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab201015 at 1/500 dilution followed by ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/1000 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 1/1000 dilution.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (ab201015)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (ab201015)

    Immunohistochemical analysis of paraffin-embedded Human glioma tissue labeling ERK2 (phospho T185) with ab201015 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear with weak cytoplasm staining on Human glioma is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

  • Western blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (ab201015)
    Western blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (ab201015)
    All lanes : Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (ab201015) 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
    Lane 3 : Untreated PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 4 : PC-12 (Rat adrenal gland pheochromocytoma cell line) treated with 200 ng/ml NGF for 4 days whole cell lysate

    Lysates/proteins at 20 µg per lane.

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

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


    Exposure time: 10 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The induction conditions refer to PMID:12454035; PMID:17026715; PMID:22206868.

  • Dot Blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (ab201015)
    Dot Blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (ab201015)

    Dot blot analysis of ERK2 (phospho T185) labeled with ab201015 at 1/1000 dilution.

    Lane 1: ERK2 (pT185) phospho peptide: DHTGFLT(p)EYVATR  aa179-191 peptide.
    Lane 2: ERK2 Non-phospho peptide: DHTGFLTEYVATR  aa179-191 peptide.
    Lane 3: ERK2 (pY187) phospho peptide: DHTGFLTEY(p)VATR  aa179-191 peptide.

    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated (ab97051)  at 1/100000 dilution was used as secondary antibody.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: 3 minutes.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (ab201015)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (ab201015)

    Immunohistochemical analysis of paraffin-embedded hHman breast tissue labeling ERK2 (phospho T185) with ab201015 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on Human breast is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

  • Immunoprecipitation - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (ab201015)
    Immunoprecipitation - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (ab201015)

    ERK2 (phospho T185) was immunoprecipitated from 0.35 mg of NIH/3T3 (Mouse embryonic fibroblast cell line) treated with 50ng/ml PDGF for 40min whole cell lysate with ab201015 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab201015 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/1000 dilution.

    Lane 1: NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate, 10µg (Input).

    Lane 2: ab201015 IP in NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate.

    Lane 3: Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) instead of ab201015 in NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate.

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

    Exposure time: 3 minutes.

  • Immunoprecipitation - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (ab201015)
    Immunoprecipitation - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (ab201015)

    ERK2 (phospho T185) was immunoprecipitated from 0.35 mg of PC-12 (Rat adrenal gland pheochromocytoma cell line) treated with 100ng/ml NGF for 10min whole cell lysate with ab201015 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab201015 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/1000 dilution.

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

    Lane 2: ab201015 IP in PC-12 treated with 100ng/ml NGF for 10min whole cell lysate.

    Lane 3: Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) instead of ab201015 in PC-12 treated with 100ng/ml NGF for 10min whole cell lysate

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

    Exposure time: 3 minutes.

References

This product has been referenced in:
  • Chang L  et al. miRNA-126 enhances viability, colony formation, and migration of keratinocytes HaCaT cells by regulating PI3?K/AKT signaling pathway. Cell Biol Int 43:182-191 (2019). Read more (PubMed: 30571843) »
  • Altshuler A  et al. RAS Regulates the Transition from Naive to Primed Pluripotent Stem Cells. Stem Cell Reports 10:1088-1101 (2018). WB ; Mouse . Read more (PubMed: 29456180) »
See all 11 Publications for this product

Publishing research using ab201015? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-3 of 3 Abreviews or Q&A

Immunocytochemistry/ Immunofluorescence abreview for Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401]

 Good
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Vascular smooth muscle cell)
Permeabilization
Yes - NP40
Specification
Vascular smooth muscle cell
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Fixative
Formaldehyde
Read More

Abcam user community

Verified customer

Submitted Nov 06 2018

Western blot abreview for Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Vascular smooth muscle cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Treatment
2 uM U0126 MEK inhibitor (2 hours)
Specification
Vascular smooth muscle cells
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Read More

Abcam user community

Verified customer

Submitted Mar 12 2018

Immunocytochemistry/ Immunofluorescence abreview for Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401]

 Average
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Permeabilization
Yes - 0.5% Triton X-100
Specification
HeLa
Fixative
Paraformaldehyde
Read More

Dr. Kirk Mcmanus

Verified customer

Submitted Jul 26 2017

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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