| 货号 | 9925T |
| 描述 | Phospho-Syk Sampler Kit provides an economical means to evaluate the activation status of Syk, including the phosphorylation of Tyr323, Tyr352 and Tyr525/526. The control Syk Antibody is also included. The kit contains enough primary and secondary antibodies for four Western blot experiments. |
| 目标/特异性 | Phospho-Syk (Ser323) and Phospho-Syk (Tyr525/526) Antibodies detect transfected, Tyr323 and Tyr 525/526 phosphorylated human Syk. Syk Antibody and Phospho-Zap-70 (Tyr319)/Syk (Tyr352) Antibodies detect endogenous levels of total human Syk and Tyr352 phosphorylated Syk, respectively. Each phospho-Syk antibody recognizes only the specific phosphorylated form of Syk. The control Syk Antibody recognizes both nonphosphorylated and phosphorylated forms of Syk. Phospho-Zap-70 (Tyr319)/Syk (Tyr352) Antibody cross-reacts with the phosphorylated form of Zap-70. All other antibodies in the kit do not cross-react with phosphorylated or nonphosphorylated forms of other related family members. |
| 供应商 | CST |
| 背景 | Syk is a protein tyrosine kinase that plays an important role in intracellular signal transduction in hematopoietic cells (1-3). Syk interacts with immunoreceptor tyrosine-based activation motifs (ITAMs) located in the cytoplasmic domains of immune receptors (4). It couples the activated immunoreceptors to downstream signaling events that mediate diverse cellular responses, including proliferation, differentiation and phagocytosis (4). There is also evidence of a role for Syk in nonimmune cells, and Syk is a potential tumor suppressor in human breast carcinomas (5). Tyr323 is a negative regulatory phosphorylation site within the SH2-kinase linker region in Syk. Phosphorylation of Tyr323 provides a direct binding site to the TKB domain of Cbl (6,7). Tyrosine 352 of Syk is involved in the association of PLC-γ1 (8). Tyrosines 525 and 526 are located in the activation loop of the Syk kinase domain, and phosphorylation of Tyr525/526 of human Syk (equivalent to the Tyr519/520 of mouse Syk) is essential for Syk function (9). |
| 存放说明 | -20C |
| 参考文献 | Cheng, A.M. and Chan, A.C. (1997) Curr Opin Immunol 9, 528-33. Kurosaki, T. (1997) Curr Opin Immunol 9, 309-18. Chu, D.H. et al. (1998) Immunol Rev 165, 167-80. Turner, M. et al. (2000) Immunol Today 21, 148-54. Coopman, P.J. et al. (2000) Nature 406, 742-7. Deckert, M. et al. (1998) J Biol Chem 273, 8867-74. Rao, N. et al. (2001) EMBO J 20, 7085-95. Law, C.L. et al. (1996) Mol Cell Biol 16, 1305-15. Zhang, J. et al. (2000) J Biol Chem 275, 35442-7. |
Western blot analysis of extracts from various cell lines using Syk (D3Z1E) XP® Rabbit mAb. | |
Immunoprecipitation of Syk protein from SR cell extracts, using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Syk(D3Z1E) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Syk (D3Z1E) XP® Rabbit mAb | |
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Syk (D3Z1E) XP®Rabbit mAb. | |
Flow cytometric analysis of RL-7 cells using Syk (D3Z1E) XP® Rabbit mAb (blue) compared to Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red). Anti-rabbit IgG (H+L), F(ab)2 Fragment (Alexa Fluor®647 Conjugate) #4414 was used as a secondary antibody. | |
Immunohistochemical analysis of paraffin-embedded human lymph node using Syk (D3Z1E) XP®Rabbit mAb. | |
Immunohistochemical analysis of paraffin-embedded mouse spleen using Syk (D3Z1E) XP®Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right). | |
Confocal immunofluorescent analysis of Ramos cells, serum-starved (overnight; left) or IgM-treated (12 ug/ml, 2 minutes; right), using Phospho-Syk (Tyr525/526) (C87C1) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). | |
Flow cytometric analysis of Ramos cells, untreated (blue) or treated with anti-IgM (green), using Phospho-Syk(Tyr525/526) (C87C1) Rabbit mAb. Anti-rabbit IgG (H+L), F(ab)2 Fragment (PE Conjugate) #8885 was used as a secondary antibody. | |
Western blot analysis of extracts from Ramos cells, untreated or treated with anti-human-IgM (10 µg/ml), using Phospho-Syk (Tyr323) Antibody #2715 (upper) or Syk Antibody #2712 (lower). | |
Western blot analysis of extracts from Ramos cells, untreated or treated with anti-IgM, using Phospho-Syk (Tyr525/526) (C87C1) Rabbit mAb (upper) or Syk Antibody #2712 (lower). | |
Immunofluorescent analysis of Jurkat cells, CD3-treated (left) or untreated (right), using Phospho-Zap-70 (Tyr319)/Syk (Tyr352) Antibody (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye). | |
Western blot analysis of extracts from Ramos cells, untreated or treated with anti-IgM (12 µg/ ml for 2 minutes), hydrogen peroxide (10 mM for 2 minutes) or lambda phosphatase, using Phospho-Zap-70 (Tyr319)/ Syk (Tyr352) Antibody. | |
Flow cytometric analysis of Jurkat cells, untreated (blue) or CD3 treated (green), using Phospho-Zap70 (Tyr319)/Syk (Tyr352) Antibody compared to a nonspecific negative control antibody (red). | |
Two-color flow cytometric analysis of Jurkat cells, untreated (left) or anti-CD3 activated (right), using Phospho-Zap-70 (Tyr319)/Syk (Tyr352) Antibody and Phospho-p44/42 MAPK (Thr202/Tyr204) (E10) mAb #9106. Anti-CD3 activation increases the intensity of label with both antibodies. | |
Western blot analysis of extracts from 293T cells expressing recombinant wild-type or mutant Syk proteins, cotransfected with CD8, using Phospho-Syk (Tyr323) Antibody (upper) or Syk Antibody #2712 (lower). (Provided by Dr. Alagarsamy L. Reddi, laboratory of Dr. Hamid Band, Harvard University, Massachusetts.) |
