Flow cytometric analysis of Jurkat cells treated with U0126 #9903 (blue) or TPA #4174 (green) using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (197G2) Rabbit mAb (Alexa Fluor® 647 Conjugate) #13148.
The MAPK Family Alexa Fluor® 647 Conjugated Antibody Sampler Kit provides an economical means to study the activation status of members of the MAPK family of proteins without the need for a fluorescent secondary antibody.
Phospho-p38 MAPK (Thr180/Tyr182) (28B10) Mouse mAb (Alexa Fluor® 647 Conjugate) recognizes endogenous levels of p38 MAP kinase only when dually phosphorylated at Thr180 and Tyr182. This antibody does not appreciably cross-react with the corresponding phosphorylated forms of either p44/42 MAPK (Erk1/2) or SAPK/JNK. Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) recognizes endogenous levels of p44 and p42 MAPK (Erk1/2) when either dually phosphorylated at Thr202 and Tyr204 of Erk1 (Thr185 and Tyr187 of Erk2), or singly phosphorylated at Thr202. This antibody does not cross-react with the corresponding phosphorylated residues of either SAPK/JNK or p38 MAP kinases. p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb (Alexa Fluor® 647 Conjugate) recognizes endogenous levels of total p44/42 MAPK (Erk1/2) protein. This antibody does not cross-react with SAPK/JNK or p38 MAP kinases. Phospho-SAPK/JNK (Thr183/Tyr185) (G9) Mouse mAb (Alexa Fluor® 647 Conjugate) recognizes endogenous levels of p46 and p54 SAPK/JNK dually phosphorylated at Thr183 and Tyr185. This antibody does not recognize endogenous levels of phosphorylated p44/42 or p38 MAP kinases.
Monoclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr180/Tyr182 of human p38 MAP kinase protein, a synthetic phosphopeptide corresponding to residues surrounding Thr202/Tyr204 of human p44 MAP kinase protein, a synthetic peptide corresponding to residues near the carboxy terminus of rat p44 MAP kinase protein, or a synthetic phosphopeptide corresponding to residues around Thr183/Tyr185 of human SAPK/JNK protein.
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. p38 MAP kinase (MAPK), also called RK (1) or CSBP (2), is the mammalian orthologue of the yeast HOG kinase that participates in a signaling cascade controlling cellular responses to cytokines and stress (1-4). 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 (5-7), and research investigators consider it an important target in the diagnosis and treatment of cancer (8). The stress-activated protein kinase/Jun-amino-terminal kinase (SAPK/JNK) is potently and preferentially activated by a variety of environmental stresses including UV and gamma radiation, ceramides, inflammatory cytokines, and in some instances, growth factors and GPCR agonists (9).
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