Revision 7

#9911

Store at -20C

Phospho-Erk1/2 Pathway Antibody Sampler Kit

1 Kit

(5 x 20 microliters)

CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

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

Product Includes Product # Quantity Mol. Wt Isotype/Source
Phospho-c-Raf (Ser338) (56A6) Rabbit Monoclonal Antibody942720 µl74 kDaRabbit IgG
Phospho-MEK1/2 (Ser217/221) (41G9) Rabbit Monoclonal Antibody915420 µl45 kDaRabbit IgG
Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) Rabbit Monoclonal Antibody437020 µl44, 42 kDaRabbit IgG
Phospho-p90RSK (Ser380) (D3H11) Rabbit Monoclonal Antibody1198920 µl90 kDaRabbit IgG
Phospho-MSK1 (Thr581) Antibody959520 µl90 kDaRabbit
Anti-rabbit IgG, HRP-linked Antibody7074100 µlGoat

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

Description

The Phospho-Erk1/2 Pathway Antibody Sampler Kit provides an economical means of evaluating multiple members of the Erk pathway as well as their activation state. The kit contains enough primary and secondary antibodies to perform two Western blot experiments.

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.

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 7

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines, starved overnight and either untreated (-) or treated (+) with TPA #4174 (200 nM, 15 min), Human Epidermal Growth Factor (hEGF; 100 ng/mL, 15 min), or Human Platelet-Derived Growth Factor BB (hPDGF-BB; 100 ng/mL, 15 min) as indicated, using Phospho-p90RSK (Ser380) (D3H11) Rabbit mAb.
undefined Image 1: Phospho-Erk1/2 Pathway Antibody Sampler Kit
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 2: Phospho-Erk1/2 Pathway Antibody Sampler Kit
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 3: Phospho-Erk1/2 Pathway Antibody Sampler Kit
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 7

Cell Signaling Technology Logo
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
undefined Image 4: Phospho-Erk1/2 Pathway Antibody Sampler Kit
Western blot analysis of extracts from untreated or TPA-treated HeLa and NIH/3T3 cells, using Phospho-MEK1/2 (Ser217/221) (41G9) Rabbit mAb (upper), or MEK1/2 Antibody #9122 (lower).
undefined Image 5: Phospho-Erk1/2 Pathway Antibody Sampler Kit
Western blot analysis of extracts from NIH3T3, HeLa and COS cells, untreated or treated with TPA, using Phospho-c-Raf (Ser338) (56A6) Rabbit mAb.
undefined Image 6: Phospho-Erk1/2 Pathway Antibody Sampler Kit
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 7

Cell Signaling Technology Logo
Western blot analysis of extracts from 293 cells, transfected with wt MSK1 or mutant MSK1, untreated or TPA-treated (200 nM), using Phospho-MSK1 (Thr581) Antibody.
undefined Image 7: Phospho-Erk1/2 Pathway Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human colon carcinoma, control (left) or λ phosphatase-treated (right), using Phospho-p90RSK (Ser380) (D3H11) Rabbit mAb.
undefined Image 8: Phospho-Erk1/2 Pathway Antibody Sampler Kit
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 9: Phospho-Erk1/2 Pathway Antibody Sampler Kit
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 7

Cell Signaling Technology Logo
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 10: Phospho-Erk1/2 Pathway Antibody Sampler Kit
Western blot analysis of extracts from 293 cells, untreated, UV-treated or calyculin A-treated, using Phospho-MSK1 (Thr581) Antibody.
undefined Image 11: Phospho-Erk1/2 Pathway Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using Phospho-p90RSK (Ser380) (D3H11) Rabbit mAb.
undefined Image 12: Phospho-Erk1/2 Pathway Antibody Sampler Kit
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 7

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 13: Phospho-Erk1/2 Pathway Antibody Sampler Kit
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 14: Phospho-Erk1/2 Pathway Antibody Sampler Kit
Western blot analysis of extracts from 293 cells untreated or treated with UV and HeLa cells untreated or treated with calyculin A, using Phospho-MSK1 (Thr581) Antibody.
undefined Image 15: Phospho-Erk1/2 Pathway Antibody Sampler Kit
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 7

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded HeLa cell pellets, untreated (left) or treated with TPA #4174 (right), using Phospho-p90RSK (Ser380) (D3H11) Rabbit mAb.
undefined Image 16: Phospho-Erk1/2 Pathway Antibody Sampler Kit
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 17: Phospho-Erk1/2 Pathway Antibody Sampler Kit
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 18: Phospho-Erk1/2 Pathway Antibody Sampler Kit
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 7

Cell Signaling Technology Logo
Confocal immunofluorescent analysis of NIH/3T3 cells, serum-starved (left), treated with TPA #4174 (200 nM, 15 min; center), or treated with TPA followed by λ phosphatase (right), using Phospho-p90RSK (Ser380) (D3H11) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
undefined Image 19: Phospho-Erk1/2 Pathway Antibody Sampler Kit
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 20: Phospho-Erk1/2 Pathway Antibody Sampler Kit
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 21: Phospho-Erk1/2 Pathway Antibody Sampler Kit
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 7

Cell Signaling Technology Logo
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 22: Phospho-Erk1/2 Pathway Antibody Sampler Kit
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 23: Phospho-Erk1/2 Pathway Antibody Sampler Kit
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 24: Phospho-Erk1/2 Pathway Antibody Sampler Kit
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 7

Cell Signaling Technology Logo
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 25: Phospho-Erk1/2 Pathway Antibody Sampler Kit
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 26: Phospho-Erk1/2 Pathway Antibody Sampler Kit
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 27: Phospho-Erk1/2 Pathway Antibody Sampler Kit
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 7

Cell Signaling Technology Logo
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 28: Phospho-Erk1/2 Pathway Antibody Sampler Kit
Simple WesternTM analysis of lysates (0.1 mg/ml) from 3T3 + TPA cells using Phospho-MEK1/2 (Ser217/221) (41G9) Rabbit mAb #9154. 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 JessTM Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230kDa separation module.
undefined Image 29: Phospho-Erk1/2 Pathway Antibody Sampler Kit
Simple WesternTM analysis of lysates (1 mg/ml) from HeLa cells using Phospho-c-Raf (Ser338) (56A6) Rabbit mAb #9427. The virtual lane view (left) shows the target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the JessTM Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230kDa separation module.
undefined Image 30: Phospho-Erk1/2 Pathway Antibody Sampler Kit
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.