Name: Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401]
Brand: RabMAb
SKU: ab201015
Rating: 4 (4 reviews)

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

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR19401] to ERK1 (phospho T202) + ERK2 (phospho T185)
Suitable for: ICC/IF, IP, IHC-P, WB, Dot blot
Reacts with: Mouse, Rat, Human

Carrier Free

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. This information is proprietary to Abcam and/or its suppliers.
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
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.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 0.05% BSA, 40% Glycerol
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19401
Isotype

IgG
Research areas

Alternative Versions
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)

The Abpromise guarantee

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	(3)	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)	1/1000. Detects a band of approximately 44, 42 kDa (predicted molecular weight: 41 kDa).
Dot blot		1/1000.

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.

Cellular localization

ERK2: Nucleus.
Database links
- Entrez Gene: 5594 Human
- Entrez Gene: 5595 Human
- Entrez Gene: 26413 Mouse
- Entrez Gene: 26417 Mouse
- Entrez Gene: 116590 Rat
- Entrez Gene: 50689 Rat
- Omim: 176948 Human
- Omim: 601795 Human
- SwissProt: P27361 Human
- SwissProt: P28482 Human
- SwissProt: P63085 Mouse
- SwissProt: Q63844 Mouse
- SwissProt: P21708 Rat
- SwissProt: P63086 Rat
- Unigene: 431850 Human
- Unigene: 861 Human
- Unigene: 196581 Mouse
- Unigene: 8385 Mouse
- Unigene: 2592 Rat
- Unigene: 34914 Rat
see all

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)

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)

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.

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

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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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 Detection Reaction (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.
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 Detection Reaction (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.
Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (ab201015)

Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

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Datasheet download

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

ab201015 has been referenced in 107 publications.

Zhang Z et al. Batroxobin inhibits astrocyte activation following nigrostriatal pathway injury. Neural Regen Res 16:721-726 (2021). PubMed: 33063734
Wang X et al. C-C chemokine receptor 5 signaling contributes to cardiac remodeling and dysfunction under pressure overload. Mol Med Rep 23:N/A (2021). PubMed: 33200795
Zhao J et al. lncRNA LIFR-AS1 inhibits gastric carcinoma cell proliferation, migration and invasion by sponging miR-4698. Mol Med Rep 23:N/A (2021). PubMed: 33355363
Qi X et al. Saikosaponin a contributed to CCIN treatment by promoting neutrophil bactericidal activity via activation CBL-dependent ERK pathway. Phytomedicine 82:153444 (2021). PubMed: 33421903
Kadir ND et al. Electrospun fibers enhanced the paracrine signaling of mesenchymal stem cells for cartilage regeneration. Stem Cell Res Ther 12:100 (2021). PubMed: 33536060

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1-4 of 4 Abreviews or Q&A

Application

Immunocytochemistry/ Immunofluorescence

Sample

Mouse Cell (MC3T3 cells)

Permeabilization

Yes - NP40

Specification

MC3T3 cells

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C

Fixative

Formaldehyde

Abcam user community

Verified customer

Submitted Jul 17 2019

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

Abcam user community

Verified customer

Submitted Nov 06 2018

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

Abcam user community

Verified customer

Submitted Mar 12 2018

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (HeLa)

Permeabilization

Yes - 0.5% Triton X-100

Specification

HeLa

Fixative

Paraformaldehyde

DR. Kirk Mcmanus

Verified customer

Submitted Jul 26 2017

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Related Products

Applications

The Abpromise guarantee

Target

Cellular localization

Database links

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (107)

Customer reviews and Q&As

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

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

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

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