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9100
PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Kit
Primary Antibodies
PhosphoPlus Antibody Kit

PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Kit #9100

Citations (2)
PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Kit: Image 1
Western blot analysis of extracts from COS cells, untreated or treated with either U0126 #9903 (10 µM for 1h) or TPA #4174 (200 nM for 10 m), using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 and p44/42 MAPK (Erk1/2) (3A7) Mouse mAb #9107.
PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Kit: Image 2
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).
PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Kit: Image 3
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb.
PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Kit: Image 4
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.
PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Kit: Image 5
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).
PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Kit: Image 6
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).
PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Kit: Image 7
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).
PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Kit: Image 8
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.
PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Kit: Image 9
Western blot analysis of extracts from HeLa, NIH/3T3 and C6 cells, using p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb.
PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Kit: Image 10
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.
PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Kit: Image 11
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing cytoplasmic and nuclear localization, using p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb.
PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Kit: Image 12
Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb.
PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Kit: Image 13
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).
PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Kit: Image 14
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).
PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Kit: Image 15
Flow cytometric analysis of Jurkat cells, U0126-treated (blue) or PMA-treated (green), using p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb compared to a nonspecific negative control antibody (red).
PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Kit: Image 16
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.
PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Kit: Image 17
Western blot analysis of p44/42 MAPK (Erk1/2) Control Cell Extracts, treated with either U0126 or TPA, using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody #9101 (left) or p44/42 MAPK (Erk1/2) Antibody #9102 (right).
PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Kit: Image 18
Western blot analysis of p44/42 MAPK (Erk1/2) Control Cell Extracts, using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (E10) Mouse mAb #9106 (green) and p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 (red).
Inquiry Info.# 9100

Product Description

The PhosphoPlus® p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody Kit provides reagents and protocols for the rapid analysis of the phosphorylation status of p44 and p42 (Erk1 and Erk2). p44/42 MAPK (Erk1/2) Negative Control Cell Extracts: Total cell extracts from Jurkat cells treated with U0126 (MEK1/2 Inhibitor) #9903 at 10 μM for 1 hour, to serve as a negative control. Supplied in SDS Sample Buffer. p44/42 MAPK (Erk1/2) Positive Control Cell Extracts: Total cell extracts from Jurkat cells treated with TPA #4174 at 200 nM for 20 minutes, to serve as a positive control. Supplied in SDS Sample Buffer.

Specificity / Sensitivity

Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) Rabbit mAb #4370 detects endogenous levels of p42 and p44 MAP Kinase (Erk1 and Erk2) when dually phosphorylated at Thr202 and Tyr204 of Erk1 (Thr185 and Tyr187 of Erk2), and singly phosphorylated at Thr202. This antibody does not cross-react with the corresponding phosphorylated residues of either JNK/SAPK or p38 MAP kinases. p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 detects endogenous levels of total p44/42 MAP kinase (Erk1/Erk2) protein. The antibody does not cross-react with either JNK/SAPK or p38 MAP kinases.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr202/Tyr204 of human p44 MAP kinase or a synthetic peptide corresponding to residues near the C-terminus of rat p44 MAP kinase.

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.
  1. Roux, P.P. and Blenis, J. (2004) Microbiol Mol Biol Rev 68, 320-44.
  2. Baccarini, M. (2005) FEBS Lett 579, 3271-7.
  3. Meloche, S. and Pouysségur, J. (2007) Oncogene 26, 3227-39.
  4. Roberts, P.J. and Der, C.J. (2007) Oncogene 26, 3291-310.
  5. Rubinfeld, H. and Seger, R. (2005) Mol Biotechnol 31, 151-74.
  6. Murphy, L.O. and Blenis, J. (2006) Trends Biochem Sci 31, 268-75.
  7. Dalby, K.N. et al. (1998) J Biol Chem 273, 1496-505.
  8. Marais, R. et al. (1993) Cell 73, 381-93.
  9. Kortenjann, M. et al. (1994) Mol Cell Biol 14, 4815-24.
  10. Owens, D.M. and Keyse, S.M. (2007) Oncogene 26, 3203-13.

Pathways & Proteins

Explore pathways + proteins related to this product.

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For Research Use Only. Not For Use In Diagnostic Procedures.
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LumiGLO is a registered trademark of Kirkegaard & Perry Laboratories.