Recombinant
RabMAb

Recombinant Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] - BSA and Azide free (ab232370)

Overview

  • Product name

    Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] - BSA and Azide free
  • Description

    Rabbit monoclonal [EPR19401] to ERK1 (phospho T202) + ERK2 (phospho T185) - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human ERK2 aa 150-250 (phospho T185). The exact sequence is proprietary.
    Database link: P28482

  • Positive control

    • IHC-P: Human glioma tissue.
  • General notes

    Ab232370 is the carrier-free version of ab201015. 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 <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab232370 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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab232370 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
IP Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 44, 42 kDa (predicted molecular weight: 41 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

IHC is recommended for human only.

Dot blot Use at an assay dependent concentration.

Images

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab201015).

  • Immunohistochemical analysis of paraffin-embedded human 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.

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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 Detection Reagent (HRP) (ab131366), was used for detection 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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 Detection Reagent (HRP) (ab131366), was used for detection 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.

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab201015).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

References

ab232370 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab232370.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up