Recombinant Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)
![Western blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)](https://www.abcam.com/ps/products/214/ab214036/Images/ab214036-266406-anti-erk1-phospho-t202-erk2-phospho-t185-antibody-epr18444-western-blot.jpg "Western blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18444] to ERK1 (phospho T202) + ERK2 (phospho T185)
- Suitable for: Dot blot, IHC-P, WB, ICC/IF, IP
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] -
Description
Rabbit monoclonal [EPR18444] to ERK1 (phospho T202) + ERK2 (phospho T185) -
Host species
Rabbit -
Specificity
ab214036 does not react with a peptide containing ERK1 pY204 or ERK2 pY187 -
Tested applications
Suitable for: Dot blot, IHC-P, WB, ICC/IF, IPmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Jurkat treated with 200 ng/ml PMA for 30 minutes whole cell lysate; NIH/3T3 treated with 50 ng/ml PDGF for 40 minutes whole cell lysate; PC-12 treated with 200 ng/ml NGF for 4 days whole cell lysate. IHC-P: Human breast, placenta, breast cancer and glioma tissues; mouse kidney tissue; rat spleen tissue. ICC/IF: Jurkat cells treated with PMA treatment (200 ng/ml, 30min). IP: Jurkat treated with 200 ng/ml PMA for 30 minutes cell lysate; PC-12 treated with 200 ng/ml NGF for 4 days cell lysate.
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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
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
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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, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR18444 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab214036 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 |
|---|---|---|
| Dot blot |
Use at an assay dependent concentration.
|
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| IHC-P |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
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| WB |
1/1000. Detects a band of approximately 44, 42 kDa (predicted molecular weight: 43, 41 kDa).
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| ICC/IF |
1/100.
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| IP |
1/40.
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| Notes |
|---|
|
Dot blot
Use at an assay dependent concentration. |
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IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
|
WB
1/1000. Detects a band of approximately 44, 42 kDa (predicted molecular weight: 43, 41 kDa). |
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ICC/IF
1/100. |
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IP
1/40. |
Target
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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
see all
Images
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Western blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)All lanes : Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036) 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 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 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 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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![Immunocytochemistry/ Immunofluorescence - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)](https://www.abcam.com/ps/products/214/ab214036/Images/ab214036-266396-anti-erk1-phospho-t202-erk2-phospho-t185-antibody-epr18444-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)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) + ERK2 (phospho T185) with ab214036 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining increased after PMA treatment (200 ng/ml, 30min), and LP treatment decreased the PMA induced staining. For the “pan” antibody, the signal is unchanged after PMA treatment (200 ng/ml, 30min), and LP treatment. The nuclear counterstain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)](https://www.abcam.com/ps/products/214/ab214036/Images/ab214036-266400-anti-erk1-phospho-t202-erk2-phospho-t185-antibody-epr18444-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)Immunohistochemical analysis of paraffin-embedded human glioma tissue labeling ERK1 (phospho T202) + ERK2 (phospho T185) with ab214036 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nuclear and weak cytoplasmic staining on human glioma is observed [PMID:17487353].
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
-
![Immunoprecipitation - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)](https://www.abcam.com/ps/products/214/ab214036/Images/ab214036-266395-anti-erk1-phospho-t202-erk2-phospho-t185-antibody-epr18444-immunoprecipitation.jpg)
Immunoprecipitation - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)ERK1 (phospho T202) + ERK2 (phospho T185) was immunoprecipitated from 0.35 mg of Jurkat (Human T cell leukemia cell line from peripheral blood) treated with 200 ng/ml PMA for 30 minutes whole cell lysate with ab214036 at 1/40 dilution.
Western blot was performed from the immunoprecipitate using ab214036 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution
Lane 1: Jurkat treated with 200 ng/ml PMA for 30 minutes whole cell lysate 10µg (Input).
Lane 2: ab214036 IP in Jurkat treated with 200 ng/ml PMA for 30 minutes whole cell lysate.
Lane 3: Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab214036 in Jurkat treated with 200 ng/ml PMA for 30 minutes whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.
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![Dot Blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)](https://www.abcam.com/ps/products/214/ab214036/Images/ab214036-284693-anti-erk1-phospho-t202-erk2-phospho-t185-antibody-epr18444-dot-blot.jpg)
Dot Blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)Dot blot analysis of ERK1 (pT202) peptide (Lane 1), ERK1 (pT204) peptide (Lane 2), ERK1 (pT202 + pT204) peptide (Lane 3) and ERK1 non-phospho peptide (Lane 4) labelling ERK1 (pT202) with ab214036.
Exposure time: 3 minutes.
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![Western blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)](https://www.abcam.com/ps/products/214/ab214036/Images/ab214036-266405-anti-erk1-phospho-t202-erk2-phospho-t185-antibody-epr18444-western-blot.jpg)
Western blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)All lanes : Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036) 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
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 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: 2 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)](https://www.abcam.com/ps/products/214/ab214036/Images/ab214036-266403-anti-erk1-phospho-t202-erk2-phospho-t185-antibody-epr18444-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)Immunohistochemical analysis of paraffin-embedded human breast tissue labeling ERK1 (phospho T202) + ERK2 (phospho T185) with ab214036 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nuclear staining on human normal breast tissue is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
-
![Immunocytochemistry/ Immunofluorescence - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)](https://www.abcam.com/ps/products/214/ab214036/Images/ab214036-266397-anti-erk1-phospho-t202-erk2-phospho-t185-antibody-epr18444-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)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) + ERK2 (phospho T185) with ab214036 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining increased after PMA treatment (200 ng/ml, 30min), and LP treatment decreased the PMA induced staining.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1: ab214036 at 1/100 dilution followed by Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
-
![Immunoprecipitation - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)](https://www.abcam.com/ps/products/214/ab214036/Images/ab214036-266394-anti-erk1-phospho-t202-erk2-phospho-t185-antibody-epr18444-immunoprecipitation.jpg)
Immunoprecipitation - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)ERK1 (phospho T202) + ERK2 (phospho T185) was immunoprecipitated from 0.35 mg of PC-12 (Rat adrenal gland pheochromocytoma cell line) treated with 200 ng/ml NGF for 4 days whole cell lysate with ab214036 at 1/40 dilution.
Western blot was performed from the immunoprecipitate using ab214036 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution
Lane 1: PC-12 treated with 200 ng/ml NGF for 4 days whole cell lysate 10µg (Input).
Lane 2: ab214036 IP in PC-12 treated with 200 ng/ml NGF for 4 days whole cell lysate.
Lane 3: Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab214036 in PC-12 treated with 200 ng/ml NGF for 4 days whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.
-
![Western blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)](https://www.abcam.com/ps/products/214/ab214036/Images/ab214036-266404-anti-erk1-phospho-t202-erk2-phospho-t185-antibody-epr18444-western-blot.jpg)
Western blot - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)All lanes : Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036) at 1/1000 dilution
Lane 1 : Untreated PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma cell line) treated with 200 ng/ml NGF for 4 days whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 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 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)](https://www.abcam.com/ps/products/214/ab214036/Images/ab214036-266402-anti-erk1-phospho-t202-erk2-phospho-t185-antibody-epr18444-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling ERK1 (phospho T202) + ERK2 (phospho T185) with ab214036 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nuclear and weak cytoplasmic staining on scattered cells of human placenta is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)](https://www.abcam.com/ps/products/214/ab214036/Images/ab214036-266401-anti-erk1-phospho-t202-erk2-phospho-t185-antibody-epr18444-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling ERK1 (phospho T202) + ERK2 (phospho T185) with ab214036 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nuclear and weak cytoplasmic staining on human breast tissue cancer is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)](https://www.abcam.com/ps/products/214/ab214036/Images/ab214036-266399-anti-erk1-phospho-t202-erk2-phospho-t185-antibody-epr18444-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling ERK1 (phospho T202) + ERK2 (phospho T185) with ab214036 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nuclear and weak cytoplasmic staining on scattered cells of mouse kidney is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)](https://www.abcam.com/ps/products/214/ab214036/Images/ab214036-266398-anti-erk1-phospho-t202-erk2-phospho-t185-antibody-epr18444-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling ERK1 (phospho T202) + ERK2 (phospho T185) with ab214036 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nuclear and weak cytoplasmic staining on scattered cells of rat spleen is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
-
![Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)](https://www.abcam.com/ps/products/214/ab214036/Images/ab214036-16-benefits-of-recombinant-antibodies.png)
Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR18444] (ab214036)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (22)
ab214036 has been referenced in 22 publications.
- Swiatnicki M et al. Profiling oncogenic KRAS mutant drugs with a cell-based Lumit p-ERK immunoassay. SLAS Discov 27:249-257 (2022). PubMed: 35288294
- Zhang RZ et al. Qi Sui Zhu Shui Plaster Inhibits AQP1 and MAPK Signaling Reduces Liver Damage Induced by Cirrhotic Ascites. J Healthc Eng 2022:9928546 (2022). PubMed: 35399826
- Lv B et al. Network Pharmacology Experiments Show That Emodin Can Exert a Protective Effect on MCAO Rats by Regulating Hif-1α/VEGF-A Signaling. ACS Omega 7:22577-22593 (2022). PubMed: 35811865
- Ji W et al. Triptolide attenuates inhibition of ankylosing spondylitis-derived mesenchymal stem cells on the osteoclastogenesis through modulating exosomal transfer of circ-0110634. J Orthop Translat 36:132-144 (2022). PubMed: 36185580
- Lu H et al. Propofol suppresses cell viability, cell cycle progression and motility and induces cell apoptosis of ovarian cancer cells through suppressing MEK/ERK signaling via targeting circVPS13C/miR-145 axis. J Ovarian Res 14:30 (2021). PubMed: 33563314