Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Rat, Xenograft

Applications

Validated:

Western Blot, Intracellular Staining by Flow Cytometry, Immunocytochemistry, Simple Western, CyTOF-ready

Cited:

Immunohistochemistry-Frozen, Western Blot, Flow Cytometry, Immunocytochemistry, Simple Western, Functional Assay

Label

Unconjugated

Antibody Source

Monoclonal Rabbit IgG Clone # 269434
View all ERK1/ERK2 Primary Antibodies »

Product Specifications

Immunogen

Phosphopeptide containing ERK1 T202/Y204 site

Specificity

Detects human, mouse, and rat ERK1 and ERK2 dually phosphorylated at T202/Y204 or T185/Y187, respectively.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Human/Mouse/Rat Phospho-ERK1 (T202/Y204)/ERK2 (T185/Y187) Antibody

Detection of Mouse, Rat, and Human Phospho-ERK1 (T202/Y204) and ERK2 (T185/Y187) antibody by Western Blot.

Detection of Mouse, Rat, and Human Phospho-ERK1 (T202/Y204) and ERK2 (T185/Y187) by Western Blot.

Western blot shows lysates of NIH-3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 100 ng/mL Rabbit Anti-Human PDGF (Catalog # 120-HD) and PC-12 rat adrenal pheochromocytoma cell line untreated or treated with 100 ng/mL Recombinant Rat beta -NGF (Catalog # 556-NG) and HeLa human cervical epithelial carcinoma cell line untreated or treated with 200 nM PMA. PVDF membrane was probed with 0.5 µg/mL of Human/Mouse/Rat Phospho-ERK1 (T202/Y204)/ERK2 (T185/Y187) Monoclonal Antibody (MAB1018), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). Specific bands were detected for Phospho-ERK1 (T202/Y204)/ ERK2 (T185/Y187) at approximately 40-45 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 3.
Phospho-ERK1 (T202/Y204)/ERK2 (T185/Y187) antibody in HeLa Human Cell Line by Immunocytochemistry (ICC).

Phospho-ERK1 (T202/Y204)/ERK2 (T185/Y187) in HeLa Human Cell Line.

ERK1/ERK2 phosphorylated at T202/Y204 (ERK1) and T185/Y187 (ERK2) was detected in immersion fixed Hela human cervical epithelial carcinoma cells, unstimulated (lower panel) and stimulated (upper panel) with PMA using Rabbit Anti-Human/Mouse/Rat Phospho-ERK1/ERK2 (ERK1 T202/Y204, ERK2 T185/Y187) Monoclonal Antibody (Catalog # MAB1018) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Phospho-ERK1/ERK2 antibody in PMA-treated Jurkat Human Cell Line antibody by Flow Cytometry.

Detection of Phospho-ERK1/ERK2 in PMA-treated Jurkat Human Cell Line by Flow Cytometry.

Jurkat human acute T cell leukemia cell line were unstimulated (light orange open histogram) or treated with 50 ng/mL PMA for 10 minutes (dark orange filled histogram), then stained with Rabbit Anti-Human/Mouse/Rat Phospho-ERK1/ERK2 (ERK1 T202/Y204, ERK2 T185/Y187) Monoclonal Antibody (Catalog # MAB1018) or control antibody (Catalog # AB-105-C, blue open histogram), followed by Fluorescein-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0112). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with ice-cold methanol.
Detection of Human Phospho-ERK1 (T202/Y204)/ERK2 (T185/Y187) antibody by Simple WesternTM.

Detection of Human Phospho-ERK1 (T202/Y204)/ERK2 (T185/Y187) by Simple WesternTM.

Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line untreated (-) or treated (+) with 200 nM PMA and Ionomycin for 20 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-ERK1 (T202/Y204)/ERK2 (T185/Y187) at approximately 42 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human/Mouse/Rat Phospho-ERK1/ERK2 (ERK1 T202/Y204, ERK2 T185/Y187) Monoclonal Antibody (Catalog # MAB1018). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Detection of Human ERK1/ERK2 by Flow Cytometry

Detection of Human ERK1/ERK2 by Flow Cytometry

Differentiation Characteristics of Individual Stem Cells.(A) Flow cytometric analysis of undifferentiated stem cell markers in the clonal NTERA2.Tom cell line. Black line depicts P3X negative control and green depicts antigen specific expression (OCT4, SOX2, NANOG and SSEA4). (B) Graph depicting the percentage of TUJ1 positive neurons within differentiated colonies derived from individual RA treated NTERA2.Tom stem cells. (C) Images of differentiated NTERA2.Tom colonies derived from individual NTERA2.Tom cells on a background of wildtype NTERA2 cells. Cell were exposed to RA for 21 days (Green - beta III tubulin, Blue - Hoechst) White arrows depict beta III tubulin positive NTERA2.Tom neurons. Scale bar represents 50 µM. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/20531938), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human ERK1/ERK2 by Western Blot

Detection of Human ERK1/ERK2 by Western Blot

p38 inhibitor SB203580 improves the up-regulation of MMP-1 by DDC through stimulating ERK1/2Co-cultures were treated with or without 100 μM DDC for 1 h in the presence or absence of p38 inhibitor (SB203580, 10 μM). Phosphorylation of ERK1/2, p38 and Akt in LX-2 cells of co-cultures were determined by Western blotting analysis. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/23577625), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human ERK1/ERK2 by Western Blot

Detection of Human ERK1/ERK2 by Western Blot

Phospho-proteome profiling results of LX-2 cells co-cultured with C3A–CYP2E1 cells with or without DDC treatment and detection of the effect of DDC on intracellular kinases in LX-2 cells of co-cultures200 μg of total cell lysates from LX-2 cells co-cultured with C3A-2E1 cells with or without 100 μM DDC for 1 h were incubated with membranes of the human phospho-MAPK Array Kit according to the manufacturer's instructions. Phospho MAPK Array data were developed on X-ray films following exposure to chemiluminescent reagents. 20 μg aliquots of total cell lysates from LX-2 cells were subjected to Western blotting analysis. (A) Template showing the location of MAPK antibodies spotted onto the human phospho-MAPK Array Kit. (B) The activation status of ERK1/2 in LX-2 cells co-cultured with C3A-2E1 cells after DDC treatment. (C) The activation status of p38 in LX-2 cells co-cultured with C3A-2E1 cells after DDC treatment. (D) The activation status of Akt in LX-2 cells co-cultured with C3A-2E1 cells after DDC treatment. (E) The activation status of ERK1/2, p38 and Akt in LX-2 cells co-cultured with C3A cells after DDC treatment. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/23577625), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human ERK1/ERK2 by Western Blot

Detection of Human ERK1/ERK2 by Western Blot

DDC up-regulates MMP-1 through ERK1/2 and Akt activation(A) Co-cultures were treated with 100 μM DDC for the indicated time. Phosphorylation of ERK1/2, p38 and Akt were determined by Western blotting analysis. The corresponding non-phosphorylated ERK1/2, p38, Akt and beta -actin were used for protein loading control. (B) Co-cultures were treated with or without 100 μM DDC for 24 h in the presence or absence of ERK1/2 inhibitor (U0126, 10 μM). MMP-1 protein levels were analysed by Western blotting. (C) Co-cultures were treated with or without 100 μM DDC for 24 h in the presence or absence of p38 inhibitor (SB203580, 10 μM). MMP-1 protein levels were analysed by Western blotting. (D) Co-cultures were treated with or without 100 μM DDC for 24 h in the presence or absence of Akt inhibitor (T3830, 50 μM). MMP-1 protein levels were analysed by Western blotting. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/23577625), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human ERK1/ERK2 by Western Blot

Detection of Human ERK1/ERK2 by Western Blot

Phospho-proteome profiling results of LX-2 cells co-cultured with C3A–CYP2E1 cells with or without DDC treatment and detection of the effect of DDC on intracellular kinases in LX-2 cells of co-cultures200 μg of total cell lysates from LX-2 cells co-cultured with C3A-2E1 cells with or without 100 μM DDC for 1 h were incubated with membranes of the human phospho-MAPK Array Kit according to the manufacturer's instructions. Phospho MAPK Array data were developed on X-ray films following exposure to chemiluminescent reagents. 20 μg aliquots of total cell lysates from LX-2 cells were subjected to Western blotting analysis. (A) Template showing the location of MAPK antibodies spotted onto the human phospho-MAPK Array Kit. (B) The activation status of ERK1/2 in LX-2 cells co-cultured with C3A-2E1 cells after DDC treatment. (C) The activation status of p38 in LX-2 cells co-cultured with C3A-2E1 cells after DDC treatment. (D) The activation status of Akt in LX-2 cells co-cultured with C3A-2E1 cells after DDC treatment. (E) The activation status of ERK1/2, p38 and Akt in LX-2 cells co-cultured with C3A cells after DDC treatment. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/23577625), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human ERK1/ERK2 by Western Blot

Detection of Human ERK1/ERK2 by Western Blot

ERK, but not PI3K/Akt, mediates the increase of cyclin E induced by CREG overexpression in HUVEC. (A) Expression of p-ERK, ERK, p-Akt, Akt, cyclin E and beta -tubulin was detected by Western blotting in the three groups of cells with LY294002 (50 μM) or PD098059 (20 μM) treatment. Vehicle (DMSO)-treated cells were used as control, and beta -tubulin was used as a loading control; (B,C) Pooled analysis of the levels of Akt (B) and ERK (C) signaling activation accessed by the grayscale ratios of p-Akt/Akt and p-ERK/ERK, respectively. Data are given as the mean ± SD (n = 3). *p < 0.05; **p < 0.01. NS: no significant difference. (D) After normalization to beta -tubulin, the difference in the expression of cyclin E between the HUVEC-AdGFP and HUVEC-AdCREG group was reduced by the treatment of PD098059, but not LY294002. Data are given as the mean ± SD (n = 3). **p < 0.01. NS: no significant difference. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/24018888), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human ERK1/ERK2 by Western Blot

Detection of Human ERK1/ERK2 by Western Blot

CREG overexpression activates both ERK&PI3K/Akt signaling pathways,&ERK, but not PI3K/Akt, mediates CREG effects on HUVEC proliferation. (A) In the three groups of cells (HUVEC, AdGFP&AdCREG), expression of JNK, p38, ERK, PI3K&Akt signaling molecules&their phosphorylated (p-) forms detected by WB, with beta -tubulin as the loading control; Image collected & cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/24018888), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Phospho-ERK1 (T202/Y204)/ERK2 (T185/Y187) by Western Blot

Detection of Phospho-ERK1 (T202/Y204)/ERK2 (T185/Y187) by Western Blot

Müller cells were pre-incubated with TIMP-3 (100 ng/ml) or dilution medium for 1 h before increasing the sugar content of the cultures [25 mM of mannitol as control or 25 mM of glucose (HG)]. After 24 h levels of the p65 subunit of NF-kappa B [panel (A)], phospho-ERK1/2 [panel (B)], caspase-3 [panel (C)], and ADAM17 [panel (D)] in cell lysates was determined by Western blot analysis. Results are expressed as median (interquartile range) from three different experiments performed in triplicate. Kruskal-Wallis test and Mann-Whitney test were used for comparisons between three groups and two groups, respectively. *p < 0.05 compared with values obtained from cells treated with mannitol. #p < 0.05 compared with values obtained from cells treated with HG. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35082694), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human/Mouse/Rat Phospho-ERK1 (T202/Y204)/ERK2 (T185/Y187) Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed HeLa human cervical epithelial carcinoma cell line stimulated with PMA

Intracellular Staining by Flow Cytometry

2.5 µg/106 cells
Sample: Jurkat human acute T cell leukemia cell line treated with PMA, fixed with paraformaldehyde and permeabilized with ice-cold methanol

Simple Western

5 µg/mL
Sample: Jurkat human acute T cell leukemia cell line treated with PMA and Ionomycin

Western Blot

0.5 µg/mL
Sample: NIH‑3T3 mouse embryonic fibroblast cell line treated with Human PDGF (Catalog # 120-HD) and PC‑12 rat adrenal pheochromocytoma cell line treated with Recombinant Rat beta -NGF (Catalog # 556-NG)

Reviewed Applications

Read 2 reviews rated 5 using MAB1018 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: ERK1/ERK2

ERK1 and ERK2 (also known as MAPK3 and MAPK1) are 44- and 42-kDa Ser/Thr kinases, respectively. They are part of the Ras-Raf-ERK signal transduction cascade often found downstream of growth factor receptor activation. ERK1 and ERK2 were initially isolated and cloned as kinases activated in response to insulin and NGF. They are expressed in most, if not all, mammalian tissues. Dual threonine and tyrosine phosphorylation activate both ERKs, at Thr202/Tyr204 for human ERK1 and Thr185/Tyr187 for human ERK2. The two proteins share 83% amino acid identity, differing mainly at the N and C termini.

Long Name

Extracellular Signal-regulated Kinase 1/2

Alternate Names

ERK1, ERK-1, ERT2, Extracellular signal-regulated kinase 1, extracellular signal-related kinase 1, HS44KDAP, HUMKER1A, Insulin-stimulated MAP2 kinase, MAPK 1, Microtubule-associated protein 2 kinase, mitogen-activated protein kinase 3, p44erk1, p44-ERK1, p44mapk, p44-MAPK, PRKM3

Additional ERK1/ERK2 Products

Product Documents for Human/Mouse/Rat Phospho-ERK1 (T202/Y204)/ERK2 (T185/Y187) Antibody

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Product Specific Notices for Human/Mouse/Rat Phospho-ERK1 (T202/Y204)/ERK2 (T185/Y187) Antibody

For research use only

Citations for Human/Mouse/Rat Phospho-ERK1 (T202/Y204)/ERK2 (T185/Y187) Antibody

Customer Reviews for Human/Mouse/Rat Phospho-ERK1 (T202/Y204)/ERK2 (T185/Y187) Antibody (2)

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  • Human/Mouse/Rat Phospho-ERK1 (T202/Y204)/ERK2 (T185/Y187) Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: HeLa cells
    Species: Human
    Verified Customer | Posted 10/11/2021
    Human/Mouse/Rat Phospho-ERK1 (T202/Y204)/ERK2 (T185/Y187) Antibody MAB1018
  • Human/Mouse/Rat Phospho-ERK1 (T202/Y204)/ERK2 (T185/Y187) Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Cell Extracts
    Species: Human
    Verified Customer | Posted 06/29/2016

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