| 货号 | 9920T |
| 描述 | Phospho-(Ser/Thr) Kinase Substrate Antibody Sampler Kit contains 40 µl of each polyclonal primary antibody [Phospho-(Ser) PKC Substrate Antibody, Phospho-(Ser) CDKs Substrate Antibody and Phospho-(Ser/Thr) ATM/ATR Substrate Antibody], 40 µl each of Phospho-(Ser/Thr) Akt Monoclonal Substrate Antibody and Phospho-(Ser/Thr) PKA Substrate Antibody and 40 µl of Phospho-(Thr) MAPK/CDK Substrate mAb. |
| 目标/特异性 | Each antibody detects endogenous levels of phospho-(Ser/Thr) proteins of specific kinase substrate groups. Phospho-(Ser/Thr) Akt Substrate (110B7E) Rabbit mAb preferentially recognizes peptides and proteins containing phospho-serine/threonine preceded by arginine at positions -5 and -3, in a manner largely independent of the surrounding amino acid sequence. Some cross-reactivity is observed for peptides that contain phospho-serine/threonine preceded by arginine at position -3 . No cross-reactivity is observed with the corresponding nonphosphorylated sequences or with other phospho-serine/threonine-containing motifs. By ELISA, the antibody recognizes a wide range of peptides with phospho-threonine or Phospho-serine with arginine at -3 and -5 position. Phospho-(Ser) PKC Substrate Antibody detects endogenous levels of many cellular proteins only when phosphorylated at serine residues surrounded by Arg or Lys at the -2 and +2 positions and a hydrophobic residue at the +1 position. The antibody does not cross-react with nonphosphorylated serine residues, with phospho-threonine in the same motif, or with phospho-serine in other motifs. Phospho-PKA Substrate (RRXS*/T*) (100G7E) Rabbit mAb detects peptides and proteins containing a phospho-Ser/Thr residue with arginine at the -3 and -2 positions. It is a useful tool in identifying new substrates of PKA. The antibody recognizes other -3 arginine-bearing phospho-Ser/Thr peptides, such as substrate motifs for Akt and PKC, to a lesser extent. It does not recognize the nonphosphorylated substrate motif peptides. Phospho-(Thr) MAPK/CDK Substrate Mouse mAb detects phospho-threonine only when followed by proline. It reacts with proteins/peptides phosphorylated on the Thr-Pro motif in an otherwise highly context-independent fashion. The antibody does not cross-react with phospho-threonine in the absence of an adjacent proline. The antibody does not cross-react with phospho-tyrosine, but does react with some phospho-serine peptides containing the phospho-serine-proline motif (e.g., phospho-Elk-1). Phospho-(Ser) CDKs Substrate Antibody detects phospho-serine in a (K/R)(S*)PX(K/R) motif. The antibody is phospho-specific but does not recognize phospho-serine in the absence of the CDK motif. The antibody does not cross-react with phospho-threonine- or phospho-tyrosine-containing peptides/proteins. Phospho-(Ser/Thr) ATM/ATR Substrate Antibody detects endogenous levels of proteins containing the ATM/ATR substrate motif. This antibody preferentially binds peptides and proteins that contain phospho-Ser/Thr preceded by Leu or similar hydrophobic amino acids at the -1 position and followed by Gln at the +1 position. The antibody does not cross-react with corresponding nonphosphorylated sequences or with other phospho-Ser/Thr-containing motifs. |
| 供应商 | CST |
| 背景 | Phospho-(Ser/Thr) kinases and phosphatases play critical roles in a wide range of biological processes. Each phospho-(Ser/Thr) kinase phosphorylates serine or threonine within a specific motif. Akt phosphorylates substrates at a serine or threonine only in a conserved motif characterized by arginine at positions -5 and -3 (1). Conventional PKC isozymes phosphorylate substrates containing serine or threonine, with arginine or lysine at the -3, -2 and +2 positions, and a hydrophobic amino acid at position +1 (2,3). A consensus phosphorylation site of PKA is serine or threonine with arginine at the -2 and -3 positions (3). The MAPK and CDK families of serine/threonine protein kinases phosphorylate threonine or serine followed by proline residue (3-5).The consensus amino acid sequence for CDK substrate is (K/R)(S*)PX(K/R), where denotes any one of the 20 amino acids and S* is the phosphorylation site (6-8). ATM and the related kinase ATR phosphorylate serine or threonine in an S*/T*Q motif (9,10). Antibodies specific to particular kinase substrates are invaluable reagents in determining kinase activity and identifying potential new kinase substrates. CST has developed antibodies that recognize phosphorylated serine or threonine within the context of a protein motif that is phosphorylated by Akt, PKC, PKA, MAPK/CDK, CDKs or ATM/ATR. As shown by peptide pairing ELISA, each phospho-(Ser/Thr) kinase substrate antibody in this sampler kit is specific to its kinase substrate motif. |
| 存放说明 | -20C |
| 参考文献 | Alessi, D.R. et al. (1996) FEBS Lett 399, 333-8. Nishikawa, K. et al. (1997) J Biol Chem 272, 952-60. Pearson, R.B. and Kemp, B.E. (1991) Methods Enzymol 200, 62-81. Songyang, Z. et al. (1996) Mol Cell Biol 16, 6486-93. Songyang, Z. (1999) Prog Biophys Mol Biol 71, 359-72. Holmes, J.K. and Solomon, M.J. (1996) J Biol Chem 271, 25240-6. Kastan, M.B. and Lim, D.S. (2000) Nat Rev Mol Cell Biol 1, 179-86. Zhao, H. and Piwnica-Worms, H. (2001) Mol Cell Biol 21, 4129-39. |
Phospho-PKA Substrate (RRXS*/T*) (100G7E) Rabbit mAb #9624: Signal-to-noise ratio of phospho- versus nonphospho-peptides. (T* and S* denote phosphorylated threonine and serine.) 使用Phospho-PKA Substrate (RRXS*/T*) (100G7E) Rabbit mAb #9624进行ELISA实验：含磷酸化肽段和非磷酸化肽段的信噪比。（T*和S*代表磷酸化的苏氨酸和丝氨酸） | |
Western blot analysis of extracts from serum-starved A431 cells, phosphorylated in vitro with PKA kinase or treated in culture with forskolin/IBMX, using Phospho-PKA Substrate (RRXS*/T*) (100G7E) Rabbit mAb #9624. Lysis buffer: 1.0% Triton X-100 (lanes 1 and 2), 2.0% SDS (lanes 3 and 4). 对血清饥饿处理的A431细胞抽提液，使用PKA激酶在体外磷酸化，或forskolin/IBMX培养，使用Phospho-PKA Substrate (RRXS*/T*) (100G7E) Rabbit mAb #9624进行Western blot分析。裂解缓冲液：1.0% Triton X-100 (列1和列2), 2.0% SDS (列3和列4)。 | |
使用Phospho-(Ser/Thr) Akt Substrate (110B7E) mAb #9614 进行ELISA实验：含磷酸化肽段和非磷酸化肽段的信噪比。（T*和S*代表磷酸化的苏氨酸和丝氨酸） | |
Phospho-(Ser/Thr) Akt Substrate (110B7E) mAb #9614 ELISA assay: Signal-to-noise ratio of phospho- versus nonphospho-peptides. (T* and S* denote phosphorylated threonine and serine.) | |
Western blot analysis of extracts from serum-starved NIH/3T3 cells, phosphorylated in vitro with Akt kinase, or treated in culture with PDGF or FBS/Calyculin A, using Phospho-(Ser/Thr) Substrate (110B7E) Rabbit mAb #9614. 对血清饥饿处理的NIH/3T3细胞抽提液，使用Akt激酶在体外磷酸化，或PDGF或FBS/Calyculin A培养，使用Phospho-(Ser/Thr) Substrate (110B7E) Rabbit mAb #9614进行Western blot分析。 | |
使用Phospho-PKA Substrate (RRXS*/T*) (100G7E) Rabbit mAb兔单抗进行ELISA实验：含磷酸化肽段或者非磷酸化肽段的信噪比 (T* and S* 代表磷酸化的苏氨酸和丝氨酸。) | |
Phospho-PKA Substrate (RRXS*/T*) (100G7E) Rabbit mAb ELISA Assay: Signal-to-noise ratio of phospho- versus nonphospho-peptides. (T* and S* denote phosphorylated threonine and serine.) | |
Western blot analysis of extracts from serum-starved A431 cells, phosphorylated in vitro with PKA kinase or treated in culture with forskolin/IBMX, using Phospho-PKA Substrate (RRXS*/T*) (100G7E) Rabbit mAb. Lysis buffer: 1.0% Triton X-100 (lanes 1 and 2), 2.0% SDS (lanes 3 and 4). 对血清饥饿处理的A431细胞抽提液，使用PKA激酶在体外磷酸化，或forskolin/IBMX培养，使用Phospho-PKA Substrate (RRXS*/T*) (100G7E) Rabbit mAb进行Western blot分析。裂解缓冲液：1.0% Triton X-100 (列1和列2), 2.0% SDS (列3和列4)。 | |
Immunohistochemical analysis of paraffin-embedded renal cell carcinoma, using Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb. 免疫组化分析石蜡包埋的肾癌，使用的抗体为Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb兔单抗 | |
Immunohistochemical analysis of paraffin-embedded lung carcinoma, using Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb. 免疫组化分析石蜡包埋的肺癌，使用的抗体为Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb兔单抗 | |
使用Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb兔单抗进行ELISA实验：含磷酸化肽段或者非磷酸化肽段的信噪比 (T* and S* 代表磷酸化的苏氨酸和丝氨酸。) | |
Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb ELISA Assay: Signal-to-noise ratio of phospho- versus nonphospho-peptides. (T* and S* denote phosphorylated threonine and serine.) |
