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877-616-CELL (2355)

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877-678-TECH (8324)

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For Research Use Only. Not for Use in Diagnostic Procedures.

Product Includes Product # Quantity Mol. Wt Isotype/Source
Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) Rabbit Monoclonal Antibody4370200 µl44, 42 kDaRabbit IgG
p44/42 MAPK (Erk1/2) (137F5) Rabbit Monoclonal Antibody4695200 µl42, 44 kDaRabbit IgG

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

UniProt ID:
#P27361, #P28482

Entrez-Gene Id:
5595, 5594

Description

PhosphoPlus® Duets from Cell Signaling Technology (CST) provide a means to assess protein activation status. Each Duet contains an activation-state and total protein antibody to your target of interest. These antibodies have been selected from CST's product offering based upon superior performance in specified applications.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Background

Mitogen-activated protein kinases (MAPKs) are a widely conserved family of serine/threonine protein kinases involved in many cellular programs, such as cell proliferation, differentiation, motility, and death. The p44/42 MAPK (Erk1/2) signaling pathway can be activated in response to a diverse range of extracellular stimuli, including mitogens, growth factors, and cytokines (1-3), and research investigators consider it an important target in the diagnosis and treatment of cancer (4). Upon stimulation, a sequential three-part protein kinase cascade is initiated, consisting of a MAP kinase kinase kinase (MAPKKK or MAP3K), a MAP kinase kinase (MAPKK or MAP2K), and a MAP kinase (MAPK). Multiple p44/42 MAP3Ks have been identified, including members of the Raf family, as well as Mos and Tpl2/COT. MEK1 and MEK2 are the primary MAPKKs in this pathway (5,6). MEK1 and MEK2 activate p44 and p42 through phosphorylation of activation loop residues Thr202/Tyr204 and Thr185/Tyr187, respectively. Several downstream targets of p44/42 have been identified, including p90RSK (7) and the transcription factor Elk-1 (8,9). p44/42 are negatively regulated by a family of dual-specificity (Thr/Tyr) MAPK phosphatases, known as DUSPs or MKPs (10), along with MEK inhibitors, such as U0126 and PD98059.

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

PhosphoPlus is a registered trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human urothelial carcinoma using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb performed on the Leica BOND RX.
undefined Image 1: PhosphoPlus<sup>®</sup> p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet
Western blot analysis of extracts from various cell lines treated with TPA (200nM, 4 hr) or PDGF (100ng/mL, 10 min) as indicated, using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb (upper), p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).
undefined Image 2: PhosphoPlus<sup>®</sup> p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet
Western blot analysis of extracts from various cell lines using p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb.
undefined Image 3: PhosphoPlus<sup>®</sup> p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Western blot analysis of extracts from 293, NIH/3T3 and C6 cells, treated with λ phosphatase or TPA #4174 as indicated, using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb (upper), or p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 (lower).
undefined Image 4: PhosphoPlus<sup>®</sup> p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet
Immunohistochemical analysis of paraffin-embedded human squamous cell carcinoma of the skin using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb performed on the Leica BOND RX.
undefined Image 5: PhosphoPlus<sup>®</sup> p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet
Western blot analysis of extracts from Hek 293 cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® p44/42 MAPK (Erk1/2) siRNA (+), using p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 and α-Tubulin (11H10) Rabbit mAb #2125. The p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb confirms silencing of p44/42 expression and α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of p44/42 MAPK (Erk1/2) siRNA.
undefined Image 6: PhosphoPlus<sup>®</sup> p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb.
undefined Image 7: PhosphoPlus<sup>®</sup> p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb performed on the Leica BOND RX.
undefined Image 8: PhosphoPlus<sup>®</sup> p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing cytoplasmic and nuclear localization, using p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb.
undefined Image 9: PhosphoPlus<sup>®</sup> p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human lung carcinoma, untreated (left) or λ phosphatase-treated (right), using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb.
undefined Image 10: PhosphoPlus<sup>®</sup> p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet
Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb performed on the Leica BOND RX.
undefined Image 11: PhosphoPlus<sup>®</sup> p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet
Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb.
undefined Image 12: PhosphoPlus<sup>®</sup> p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Immunohistochemical analysis using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb on SignalSlide™ Phospho-p44/42 MAPK (Thr202/Tyr204) IHC Controls #8103 (paraffin-embedded NIH/3T3 cells, treated with U0126 #9903 (left) or TPA #4174 (right).
undefined Image 13: PhosphoPlus<sup>®</sup> p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb in the presence of control peptide (left) or #1240 p44/42 MAPK (Erk1/2) Blocking Peptide (#4695 Specific) (right).
undefined Image 14: PhosphoPlus<sup>®</sup> p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet
Confocal immunofluorescent analysis of Drosophila egg chambers, untreated (top) or λ phosphatase-treated (bottom), using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 (green) and S6 Ribosomal Protein (54D2) Mouse mAb #2317 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
undefined Image 15: PhosphoPlus<sup>®</sup> p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Confocal immunofluorescent analysis of NIH/3T3 cells, treated with either U0126 (MEK1/2 Inhibitor) #9903 (left) or PDGF (right), using p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).
undefined Image 16: PhosphoPlus<sup>®</sup> p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet
Confocal immunofluorescent analysis of HT1080 cells, starved overnight then treated with U0126 #9903 (10 uM, 2 h; left) or PDBu (Phorbol 12,13-Dibutyrate) #12808 (100 nM, 15 m; right) using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
undefined Image 17: PhosphoPlus<sup>®</sup> p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet
Immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb.
undefined Image 18: PhosphoPlus<sup>®</sup> p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Flow cytometric analysis of Jurkat cells, treated with U0126 (10 µM, 2 hrs; green) or treated with TPA #4174 (200 nM, 30 min; blue) using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
undefined Image 19: PhosphoPlus<sup>®</sup> p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet
Immunohistochemical analysis of paraffin-embedded 4T1 syngeneic tumor using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb.
undefined Image 20: PhosphoPlus<sup>®</sup> p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet
Flow cytometric analysis of Jurkat cells using p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')₂ Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
undefined Image 21: PhosphoPlus<sup>®</sup> p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Immunoprecipitation of Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) from 3T3 cell extracts. Cells were treated with TPA, (200 nM, 15 min). Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb. Western blot was performed using Phosphop44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb. Mouse Anti-rabbit IgG (Light-Chain Specific) (D4W3E) mAb #45262 was used as a secondary antibody.
undefined Image 22: PhosphoPlus<sup>®</sup> p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet
Simple Western™ analysis of lysates (0.1 mg/mL) from serum-starved HeLa cells treated with TPA (400 nM, 4 hours) using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370. The virtual lane view (left) shows the target bands (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
undefined Image 23: PhosphoPlus<sup>®</sup> p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet
Western blot analysis of extracts from Jurkat cells, treated with U0126 (10 µM, 1 hour) (negative control) or treated with TPA (200 nM, 20 min) (positive control), using p44/42 MAPK (Erk1/2) (L34F12) Mouse mAb #4696 (Panel A) and Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 (Panel B). Anti-mouse IgG (H+L) (DyLight 680 Conjugate) #5470 (red) and Anti-rabbit IgG (H+L) (DyLight 800 4X PEG Conjugate) #5151 (green) were used as secondary antibodies.
undefined Image 24: PhosphoPlus<sup>®</sup> p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Immunoprecipitation of Jurkat cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb. Western blot analysis was performed using p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb. Mouse Anti-Rabbit IgG (Light-Chain Specific) (D4W3E) mAb (HRP Conjugate) #93702 was used as a secondary antibody.
undefined Image 25: PhosphoPlus<sup>®</sup> p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet
Simple Western™ analysis of lysates (0.1 mg/mL) from HeLa cells using p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ ​​​​​​​ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
undefined Image 26: PhosphoPlus<sup>®</sup> p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet
Confocal immunofluorescent analysis of fixed frozen mouse hippocampus using p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb (green) and ProLong Gold Antifade Reagent with DAPI #8961 (blue).
undefined Image 27: PhosphoPlus<sup>®</sup> p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Duet
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.