Cell Signaling Anti-ACTIVE® JNK pAb, Rabbit, (pTPpY) Product Anti-ACTIVE® JNK pAb, Rabbit, (pTPpY) For Research Use Only. Not for Use in Diagnostic Procedures. Description: Anti-ACTIVE® JNK pAb is a polyclonal antibody from rabbit serum. The antibody is affinity purified using a dually phosphorylated peptide that corresponds to the active form of the JNK enzymes. Features: • Specificity: Preferentially detects the dually phosphorylated, active form of the stress-activated protein kinase (SAPK), also known as c-Jun N-terminal kinase, JNK. • Immunogen: Dually phosphorylated Thr/Pro/Tyr region (pTPpY) derived from the catalytic core of the active form of JNK kinase, which corresponds to Thr183 and Tyr185 of the mammalian JNK2 enzyme. • Antibody Form: Affinity-purified rabbit IgG; supplied in 10mM sodium phosphate (pH 7.4), 20mM NaCl. • Value: Anti-ACTIVE® JNK pAb is available in two convenient sizes. Cat.# V7931 will generate up to 200ml of blotting solution, sufficient for 20 Western blots of 10ml each. The larger size, Cat.# V7932, will generate up to 600ml of blotting solution, sufficient for 60 Western blots of 10ml each. Storage Conditions: Store at –20°C. Protocol Anti-ACTIVE® MAPK, JNK and p38 Polyclonal Antibodies and Anti-ACTIVE® Qualified Secondary Antibody Conjugates Technical Bulletin Part# TB262 Size Cat.# Price (Fr) 40 µl V7931 804.00 120 µl V7932 1327.00 Anti-ACTIVE® MAPK pAb, Rabbit, (pTEpY) Product Anti-ACTIVE® MAPK pAb, Rabbit, (pTEpY) For Research Use Only. Not for Use in Diagnostic Procedures. Description: Anti-ACTIVE® MAPK pAb is a polyclonal rabbit antibody. The antibody is affinity purified using a dually phosphorylated peptide that corresponds to the active form of the mitogen-activated protein (MAP) kinase enzymes. Features: • Specificity: Preferentially detects the dually phosphorylated, active form of the mitogen-activated protein kinase (MAPK) enzymes (ERK1 and ERK2). • Immunogen: Dually phosphorylated Thr/Glu/Tyr region (pTEpY) derived from the catalytic core of the active form of the mitogen-activated protein kinase (MAPK) enzymes, ERK1 and ERK2, which corresponds to Thr183 and Tyr185 of the mammalian ERK2 enzyme. • Antibody Form: Affinity-purified rabbit IgG; supplied in PBS (pH 7.4). • Value: When used at the recommended 1:5,000 dilution, this product will generate 200ml of blotting solution, sufficient for 20 Western blots of 10ml each. Storage Conditions: Store at –20°C. Protocol Anti-ACTIVE® MAPK, JNK and p38 Polyclonal Antibodies and Anti-ACTIVE® Qualified Secondary Antibody Conjugates Technical Bulletin Anti-ACTIVE® p38 pAb, Rabbit, (pTGpY) Product Anti-ACTIVE® p38 pAb, Rabbit, (pTGpY) For Research Use Only. Not for Use in Diagnostic Procedures. Description: Anti-ACTIVE® p38 pAb is a polyclonal rabbit antibody. The antibody is affinity-purified using a dually phosphorylated peptide that corresponds to the active form of the p38 enzymes. Features: • Specificity: Preferentially detects the dually phosphorylated, active form of p38 kinase. • Immunogen: Dually phosphorylated Thr/Gly/Tyr region (pTGpY) derived from the catalytic core of the active form of p38 kinase, which corresponds to Thr180 and Tyr182 of the mammalian p38 enzyme. • Antibody Form: Affinity-purified rabbit IgG; supplied in PBS (pH 7.4). Immunocytochemical detection of active JNK enzyme in Drosophila pupal retina using the Anti-ACTIVE® JNK pAb. Drosophila pupal retina at 25% of pupal development were fixed in 3% paraformaldehyde in PBS. The Anti-ACTIVE® JNK pAb was diluted 1:100 in PBS containing 10% fetal bovine serum and 0.2% Triton® X-100. Samples were incubated with the primary antibody overnight at 4˚C, washed 3 times (10 minutes each) with 0.2% Triton® X-100 and then incubated with a goat anti-rabbit Cy®3 conjugate for 2 hours at 4˚C. Whole mounts were visualized with a Zeiss® Axioskop fluorescent microscope. The results illustrate the presence of dually phosphorylated, active forms of JNK in discrete structures of the fly retinal ommatidia including intense staining of the inner cone cells as well as the mechanosensory bristles and surrounding pigment cells. The pattern of staining (which was distinct from results obtained with an antibody for active p38) and the absence of staining in control experiments (data not shown) support the high specificity of the Anti-ACTIVE® JNK pAb. Image kindly provided by David T. Miller and Ross Cagan, Department of Molecular Biology and Pharmacology, Washington University School of Medicine, St. Louis, Missouri. • Value: When used at the recommended 1:2,000 dilution, this product will generate up to 200ml of blotting solution, sufficient for 20 Western blots of 10ml each. Storage Conditions: Store at –20°C. Protocol Anti-ACTIVE® MAPK, JNK and p38 Polyclonal Antibodies and Anti-ACTIVE® Qualified Secondary Antibody Conjugates Technical Bulletin Part# TB262 Size Cat.# Price (Fr) 100 µl V1211 590.00 Part# TB262 Size Cat.# Price (Fr) 40 µl V8031 623.00 8 For complete and up-to-date product information visit: www.promega.com/catalog 167 Imaging and Immunological Detection 1825TA07_7A Pagina 170
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