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| | PhosphoDetect™ STAT Antibody Sampler Kit |
|  | Cat. No. PS1021 |
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| Isotype: IgG | | Immunogen: a series of synthetic peptides corresponding to amino acids surrounding the designated phosphorylation sites of STAT1, STAT2, STAT3, STAT5, and STAT6, each coupled to KLH | | Form: Liquid | | Formulation: Polyclonal anti-STAT antibodies supplied as 40 µl each in 150 mM NaCl, 10 mM sodium HEPES, 100 µg/ml BSA, 50% glycerol, pH 7.5 and 100 µl goat anti-rabbit IgG, HRP-conjugated in 150 mM NaCl, 10 mM sodium HEPES, 100 µg/ml BSA, 50% glycerol, pH 7.5. | | Preservative: None | | Positive Control: Phospho-STAT1: IFN-a-treated HeLa cells Phospho-STAT2: IFN-a-treated HeLa cells Phospho-STAT3: EGF-treated A431 cells, CNTF-treated SK-N-MC cells, human breast carcinoma, and mouse trigeminal ganglia Phospho-STAT5: GM-CSF-treated TF-1 cells and IFN-a-treated HeLa cells Phospho-STAT6: IL-4-treated Daudi cells and human breast carcinoma | | Negative Control: Phospho-STAT1: Untreated HeLa cells Phospho-STAT2: Untreated HeLa cells Phospho-STAT3: Untreated A431 cells Phospho-STAT5: Untreated TF-1 cells Phospho-STAT6: Untreated Daudi cells | | Kit Contents: PhosphoDetect™ Anti-STAT1 (pTyr701) Rabbit pAb, PhosphoDetect™ Anti-STAT2 (pTyr690) Rabbit pAb, PhosphoDetect™ Anti-STAT3 (pTyr705) Rabbit pAb, PhosphoDetect™ Anti-STAT3 (pSer727) Rabbit pAb, PhosphoDetect™ Anti-STAT5 (pTyr694) Rabbit pAb (Cat. No. 575142), PhosphoDetect™ Anti-STAT6 (pTyr641) Rabbit pAb (Cat. No. 575143), Goat Anti-Rabbit IgG HRP conjugate, and a data sheet. | | Comments: A convenient set of affinity purified antibodies useful for evaluating the activation status of various STAT proteins, including the phosphorylation of STAT1 at Tyr701, STAT2 at Tyr690, STAT3 at Tyr705/Ser727, STAT5 at Tyr694 and STAT6 at Tyr641. | | Ref.: Yokogami, K., et al. 2000. Curr. Biol. 10, 47. Bromberg, J.F. et al. 1999. Cell 98, 295. Cattaneo, E., et al. 1999. Trends Neurosci..22, 365. Dentelli, P., et al. 1999. J. Immunol. 163, 2151. Frank, D.A., et al. 1999. Mol. Med. 5, 432. Lim, C.P. and Cao, X. 1999. J. Biol. Chem. 274, 31055. Su, L., et al. 1999. J. Biol. Chem. 274, 31770. Leonard, W. J. and O'Shea, J.J. 1998.Annu. Rev. Immunol. 16, 293. Caldenhoven, E., et al. 1996. J. Biol. Chem. 271, 13221. Wen, Z., et al. 1995. Cell 82, 241. Darnell, J. E. Jr., et al. 1994. Science 264, 1415. |
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A: Detection of human STAT1, phosphorylated on Tyr701, by immunoblotting. Samples: Whole cell lysate from HeLa cells left untreated (-) or treated with IFN-a (+; 100 ng/ml). Primary antibody: PhosphoDetect™ Anti-STAT1 (pTyr701) Rabbit pAb (Component JA9176) (1:1000). Detection: chemiluminescence. B: Detection of human STAT1, phosphorylated on Tyr701, by immunocytochemical staining. Samples: HeLa cells left untreated (control) or treated with IFN-a (IFN-a). Primary antibody: PhosphoDetect™ Anti-STAT1 (pTyr701) Rabbit pAb (Component JA9176) (1:500) Detection: colorimetric.
G: Detection of human STAT5, phosphorylated on Tyr694, by immunoblotting. Samples: Whole cell lysate from TF-1 cells left untreated (0 hr) or treated with GM-CSF for the specified times. Primary antibody: PhosphoDetect™ Anti-STAT5 (pTyr694) Rabbit pAb (top panel) (Component JA9180) (1:1000) and Anti-STAT5 (total) (bottom panel). Detection: chemiluminescence. H: Detection of human STAT5, phosphorylated on Tyr694, by immunocytochemical staining. Samples: HeLa cells left untreated (control) or treated with IFN-a (IFN-a). Primary antibody: PhosphoDetect™ Anti-STAT5 (pTyr694) Rabbit pAb (Component JA9180) (1:500). Detection: colorimetric.
I: Detection of human STAT6, phosphorylated on Tyr641, by immunoblotting. Samples: Whole cell lysate from Daudi cells left untreated (0 hr) or treated with IL-4 for the specified times. Primary antibody: PhosphoDetect™ Anti-STAT6 (pTyr641) Rabbit pAb (top panel) (Component JA9181) (1:1000). Detection: chemiluminescence. J: Detection of human STAT6, phosphorylated on Tyr641, by staining paraffin sections. Sample: Breast carcinoma. Primary antibody: PhosphoDetect™ Anti-STAT6 (pTyr641) Rabbit pAb (Component JA9181) (1:50). Detection: DAB with hematoxylin counterstain.
C: Detection of human STAT2, phosphorylated on Tyr690, by immunoblotting. Samples: Whole cell lysate from HeLa cells left untreated (-) or treated with IFN-a (+; 100 ng/ml for 15 min). Primary antibody: PhosphoDetect™ Anti-STAT2 (pTyr690) Rabbit pAb (Component JA9177) (1:1000). Detection: chemiluminescence.
D: Detection of human STAT3, phosphorylated on Tyr705, by immunoblotting. Samples: Whole cell lysate from SK-N-MC cells left untreated (0 min, CNTF) or treated with CNTF for 10, 30, and 60 min and A431 cells left untreated (0 min, EGF) or treated with EGF for 5 and 15 min. Primary antibodies: PhosphoDetect™ Anti-STAT3 (pTyr705) Rabbit pAb (Component JA9178) (top panel) and Anti-STAT3 (total) (bottom panel). Detection: chemiluminescence. E: Detection of human STAT3, phosphorylated on Ser727, by immunoblotting. Samples: Whole cell lysate from A431 cells left untreated (0 min) or treated with EGF for 5, 30, and 60 min. Primary antibodies: PhosphoDetect™ Anti-STAT3 (pSer727) Rabbit pAb (top panel) (Component JA9179) (1:1000) and Anti-STAT3 (total) (bottom panel). Detection: chemiluminescence. F: Detection of human STAT3, phosphorylated on Ser727, by staining paraffin sections. Sample: Breast carcinoma. Primary antibody: PhosphoDetect™ Anti-STAT3 (pSer727) Rabbit pAb (Component JA9179) (1:50). Detection: DAB with hematoxylin counterstain.
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