| 货号 | 9939T |
| 描述 | The Stat Antibody Sampler Kit provides an economical means to examine multiple Stat proteins: Stat1, Stat3, Stat5 and Stat6. The kit contains enough primary and secondary antibodies to perform four Western blot experiments. |
| 目标/特异性 | Each Stat antibody in the kit recognizes only its target protein, independent of phosphorylation state. |
| 供应商 | CST |
| 背景 | Jaks (Janus Kinases) and Stats (Signal Transducers and Activators of Transcription) are utilized by receptors for a wide variety of ligands including cytokines, hormones, growth factors and neurotransmitters. Jaks, activated via autophosphorylation following ligand-induced receptor aggregation, phosphorylate tyrosine residues on associated receptors, Stat molecules and other downstream signaling proteins (1,2). The phosphorylation of Stat proteins at conserved tyrosine residues activates SH2-mediated dimerization followed rapidly by nuclear translocation. Stat dimers bind to IRE (interferon response element) and GAS (gamma interferon-activated sequence) DNA elements, resulting in the transcriptional regulation of downstream genes (1,2). The remarkable range and specificity of responses regulated by the Stats is determined in part by the tissue-specific expression of different cytokine receptors, Jaks and Stats (2,3), and by the combinatorial coupling of various Stat members to different receptors. Serine phosphorylation in the carboxy-terminal transcriptional activation domain has been shown to regulate the function of Stat1, -2, -3, -4 and -5 (1). Phosphorylation of Stat3 at Ser727 via MAPK or mTOR pathways is required for optimal transcriptional activation in response to growth factors and cytokines including IFN-gamma and CNTF (4,5). Jak/Stat pathways also play important roles in oncogenesis, tumor progression, angiogenesis, cell motility, immune responses and stem cell differentiation (6-11). |
| 存放说明 | -20C |
| 参考文献 | Darnell Jr., J. et al. (1994) Science 264, 1415-1421. Leonard, W.J. and OShea, J.J. (1998) Annu. Rev. Immunol. 16, 293-322. Caldenhoven, E. et al. (1996) J. Biol. Chem. 271, 13221-13227. Wen, Z. et al. (1995) Cell 82, 241-250. Yokogami, K. et al. (2000) Curr. Biol. 10, 47-50. Lim, C.P. and Cao, X. (1999) J. Biol. Chem. 274, 31055-31061. Bromberg, J. F. et al. (1999) Cell 98, 295-303. Su, L. et al. (1999) J. Biol. Chem. 274, 31770-31774. Dentelli, P. et al. (1999) J. Immunol. 163, 2151-2159. Cattaneo, E. et al. (1999) Trends Neurosci. 22, 365-369. Frank, D.A. (1999) Mol. Med. 5, 432-456. |
Western blot analysis of extracts from various cell lines using Stat6 (D3H4) Rabbit mAb. | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 Ramos cells starved overnight then treated with IL-4 (100 ng/ml, 30 min) and either 10 μl of Stat6 (D3H4) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP®Human DMD Intron 2 Primers #7710, human MS4A1 promoter primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one. | |
Western blot analysis of extracts from various cell lines using Stat3 (79D7) Rabbit mAb #4904. | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 Hep G2 cells treated with IL-6 (100 ng/ml) for 30 minutes, and either 20 μl of Stat3 (79D7) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using human IRF-1 promoter primers, SimpleChIP® Human c-Fos Promoter Primers #4663, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one. | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HT-1080 cells treated with IFN-γ (50 ng/ml) for 30 minutes and either 10 μl of Stat1 Antibody or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using human IRF-1 promoter primers, SimpleChIP® Human TAP1 Promoter Primers #5148, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one. | |
Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® Stat3 siRNA I (+) or SignalSilence® Stat3 siRNA II #6582 (+), using Stat3 (79D7) Rabbit mAb #4904 and α-Tubulin (11H10) Rabbit mAb #2125. The Stat3 (79D7) Rabbit mAb confirms silencing of Stat3 expression, while the α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of Stat3 siRNA. | |
Western blot analysis of extracts from HeLa cells 48 hours following mock transfection, transfection with non-targeted (control) siRNA or transfection with Stat1 siRNA. Stat1 was detected using Stat1 Antibody #9172, and p42 was detected using p42 MAPK Antibody #9108. The Stat1 Antibody confirms silencing of Stat1 expression, and the p42 MAPK Antibody is used to control for protein loading and siRNA specificity. | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 BaF3 cells starved of IL-3 for 6 hours followed by induction with IL-3 for 45 minutes, and either 20 μl of Stat5 Antibody or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Mouse CIS Intron 1 Primers #5131, mouse SOCS-3 promoter primers, and SimpleChIP® Mouse RPL30 Intron 2 Primers #7015. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one. | |
Western blot analysis of extracts from various cell lines using the Stat5 Antibody. | |
Western blot analysis of extracts from HeLa, L929 and PC12 cells, using Stat5 (3H7) Rabbit mAb. | |
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 BaF3 cells starved of IL-3 for 6 hours followed by induction with IL-3 for 45 minutes, and either 20 μl of Stat5 Antibody or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by Real-Time PCR using SimpleChIP® Mouse CIS Intron 1 Primers #5131, mouse SOCS3 promoter primers, and SimpleChIP® Mouse RPL30 Intron 2 Primers #7015. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one. | |
Western blot analysis of extracts from various cell lines using Stat3 (79D7) Rabbit mAb. | |
Western blot analysis of extracts from various cell lines using Stat6 Antibody #9362. | |
Western blot analysis of extracts from K562 and BaF3 cell lines using Stat5 Antibody #9363. | |
Western blot analysis of extracts from SK-MEL-28 cells, untreated or IFN-alpha-treated (100 ng/ml), using Phospho-Stat1 (Tyr701) Antibody #9171 (upper) or Stat1 Antibody #9172 (lower). |
