Antibodies, Proteins, Kits and Reagents for Life Science

371 894 ₸ Product size

100 µl 371 894 ₸

Order now and get it on Tuesday March 02, 2021

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

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 & Species

Application	Species
ICC/IF	Human
IHC-P	Human
IP	Mouse Rat
WB	Mouse Rat Human

See all applications and species data

Immunogen

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.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

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

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)

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