Cell Signaling Technology

Product Pathways - Motif Antibodies

Phospho-(Ser/Thr) Kinase Substrate Antibody Sampler Kit #9920

Akt   Akt1   AMPK   ATM/ATR   CDK   general   motif   phospho-threonine   PKA   PKC   substrate   threonine  

REACTIVITY

No. Size Price
9920T 1 Kit ( 6 x 20 µl ) ¥5,301.00 现货查询 购买询价
Kit Includes Quantity Applications Reactivity Homology† MW (kDa) Isotype
Phospho-AMPK Substrate Motif [LXRXX(pS/pT) MultiMab™ Rabbit mAb mix #5759 20 µl W,IP,E-P, All, Rabbit
Phospho-ATM/ATR Substrate Motif [(pS/pT) QG] MultiMab™ Rabbit mAb mix #6966 20 µl W,IP, All, Rabbit IgG
Phospho-PKC Substrate Motif [(R/KXpSX(R/K)] MultiMab™ Rabbit mAb mix #6967 20 µl W,IP,E-P, All, Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl W, Goat
Phospho-CDK Substrate Motif [(K/H)pSP] MultiMab™ Rabbit mAb mix #9477 20 µl W,IP,E-P, H,All, Rabbit IgG
Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb #9614 20 µl W,IP,IHC-P,E-P, H,M,Dm,All, Rabbit IgG
Phospho-PKA Substrate (RRXS*/T*) (100G7E) Rabbit mAb #9624 20 µl W,IP,E-P, All, Rabbit IgG

Specificity / Sensitivity

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.

各抗体识别内源性特异激酶底物中含磷酸化丝氨酸/苏氨酸的蛋白。Phospho-(Ser/Thr) Akt Substrate (110B7E) Rabbit mAb优先识别包含-5、-3位是精氨酸的磷酸化丝氨酸/苏氨酸的内源性蛋白质,与周围序列无关。此抗体与包含仅-3位上有精氨酸的磷酸化丝氨酸/苏氨酸的蛋白质有部分的交叉反应。没有发现其能与相应的非磷酸化的序列或其它基序中的磷酸化丝氨酸/苏氨酸序列发生交叉反应。ELISA中,抗体能识别很多包含-5、-3位置上有精氨酸的磷酸化丝氨酸/苏氨酸的肽段。Phospho-(Ser) PKC Substrate Antibody可识别许多内源性的包含+2和-2位上有精氨酸或赖氨酸的或+1位有疏水氨基酸的磷酸化的丝氨酸的细胞内蛋白质。该抗体不与非磷酸化丝氨酸残基、同样模体中磷酸化的苏氨酸残基、其它模体中的磷酸化丝氨酸残基发生交叉反应。Phospho-PKA Substrate (RRXS*/T*) (100G7E) Rabbit mAb识别包含精氨酸-3和-2位上有磷酸化丝氨酸/苏氨酸的肽段或蛋白质。它是识别PKA底物的有用工具。该抗体也可以以稍弱的强度识别包含-3位上有精氨酸的磷酸化丝氨酸/苏氨酸的肽段,如Akt和PKC的底物基序。它不能识别非磷酸化的底物模体蛋白。Phospho-(Thr) MAPK/CDK Substrate Mouse mAb识别被脯氨酸跟随的磷酸化苏氨酸。它与磷酸化的苏氨酸-脯氨酸基序发生反应,并不与氨基酸序列有关。此抗体与缺乏跟随脯氨酸的磷酸化苏氨酸没有交叉反应。此抗体与磷酸化酪氨酸没有交叉反应,但与一些包含磷酸化丝氨酸-脯氨酸模体的磷酸化丝氨酸蛋白(如磷酸化Elk-1)有交叉反应。Phospho-(Ser) CDKs Substrate Antibody识别(K/R)(S*)PX(K/R)模体中磷酸化的丝氨酸。此抗体是磷酸化特异性的,但是不能识别缺乏CDK模体的磷酸化丝氨酸。此抗体与包含磷酸化苏氨酸或磷酸化酪氨酸的肽段或蛋白没有交叉反应。Phospho-(Ser/Thr) ATM/ATR Substrate Antibody识别包含ATM/ATR底物模体的蛋白质。此抗体优先结合包含磷酸化丝氨酸/苏氨酸的肽段和蛋白质,还要求-1位有亮氨酸或相似的疏水氨基酸以及+1位跟随着谷氨酰胺。此抗体不与相应的非磷酸化序列或其它包含磷酸化丝氨酸/苏氨酸的基序发生交叉反应。

Source / Purification

Polyclonal antibodies are produced by immunizing animals with phosphopeptides containing the kinase substrate motif and purified by protein A and peptide affinity chromatography.Monoclonal antibody is produced by immunizing animals with synthetic phosphopeptides .Mouse monoclonal antibody (isotype: IgM) is produced by immunizing mice with a phospho-(Thr) MAPK/CDK substrate motif-containing peptide and is purified by protein A chromatography.

多克隆抗体使用包含激酶底物基序的磷酸化肽段免疫动物制备,通过蛋白A和肽段亲和色谱纯化。单克隆抗体使用合成的磷酸化肽段免疫动物制备。鼠单抗(IgM)使用包含磷酸化苏氨酸MAPK/CDK底物基序的肽段免疫动物制备,通过蛋白A色谱纯化。

Description

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.

Phospho-(Ser/Thr) Kinase Substrate Antibody Sampler Kit试剂盒包含多种40ul一抗[Phospho-(Ser) PKC Substrate Antibody, Phospho-(Ser) CDKs Substrate Antibody和Phospho-(Ser/Thr) ATM/ATR Substrate Antibody],40ul Phospho-(Ser/Thr) Akt Monoclonal Substrate Antibody和Phospho-(Ser/Thr) PKA Substrate Antibody,40ul Phospho-(Thr) MAPK/CDK Substrate mAb。

Western Blotting

Western Blotting

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO* is added and emits light during enzyme catalyzed decomposition.

一抗结合靶蛋白后,加入二抗,HRP偶联的二抗复合物形成。然后加入LumiGLO*显色发光

Western Blotting

Western Blotting

Western blot analysis of whole cell lysates of Jurkat cells untreated and treated with PMA (50 ng/ml) and ionomycin (1 µM) for 20 minutes prior to lysis. Proteins were separated by 2D electrophoresis prior to blotting and probed with Phospho-(Ser) PKC Substrate Antibody #2261.

Jurkat细胞全细胞未经处理或经50 ng/ml PMA和1 uMionomycin处理20分钟,抽提蛋白。蛋白质进行二维电泳分离,然后进行免疫印迹和杂交,使用的抗体为Phospho-(Ser) PKC Substrate Antibody #2261。

Western Blotting

Western Blotting

Western blot analysis of whole cell lysates from Jurkat cells untreated and treated with 1 µg/ml nocodazole for 12 hours prior to lysis, using Phospho-(Thr) MAPK/CDK Substrate mAb #2321. Proteins were separated by 2D electrophoresis prior to blotting.

Jurkat细胞全细胞未经处理或经1 ug/ml噻氨酯哒唑处理12小时,抽提蛋白。使用Phospho-(Thr) MAPK/CDK Substrate mAb #2321进行Western Blot分析。蛋白质进行二维电泳分离,然后进行免疫印迹实验。

ELISA-Peptide

ELISA-Peptide

Signal to noise ratio of ELISA reading, using Phospho-(Thr) MAPK/CDK Substrate mAb #2321 with phospho-Thr-Pro containing peptides (T*P) and nonphospho-Thr-Pro containing peptides. (T* denotes phosphorylated threonine.)

使用包含phospho-Thr-Pro(T*P)和nonphospho-Thr-Pro肽段的Phospho-(Thr) MAPK/CDK Substrate mAb #2321抗体进行ELISA,图示为ELISA信噪比读数。(T*代表磷酸化的苏氨酸)

Western Blotting

Western Blotting

Chk2 transfected and UV treated COS cell extracts immunoprecipitated with Chk2 antibody then detected by Western blotting using Phospho-(Ser/Thr) ATM/ATR Substrate Antibody #2851.

对Chk2转染和UV处理的COS细胞裂解液,使用Chk2抗体免疫沉淀,然后使用Phospho-(Ser/Thr) ATM/ATR Substrate Antibody #2851抗体进行Western Blot分析。

ELISA-Peptide

ELISA-Peptide

Phospho-(Ser/Thr) PKA Substrate Antibody ELISA Assay: Signal-to-noise ratio of phospho- versus nonphospho-peptides. (T* and S* denote phosphorylated threonine and serine.)

使用Phospho-(Ser/Thr) PKA Substrate Antibody进行ELISA实验: 含磷酸化肽段和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。)

ELISA-Peptide

ELISA-Peptide

Phospho-(Ser) PKC Substrate Antibody #2261 ELISA assay: Signal-to-noise ratio of phospho- versus nonphospho-peptides. (S* denotes phosphorylated serine.)

使用Phospho-(Ser) PKC Substrate Antibody #2261进行ELISA实验:磷酸化和非磷酸化的信噪比。(S* 代表磷酸化的丝氨酸)

ELISA-Peptide

ELISA-Peptide

Phospho-(Ser/Thr) ATM/ATR Substrate Antibody #2851 ELISA assay: Signal-to-noise ratio of phospho- versus nonphospho-peptides. (S* or T* denote phosphorylated serine or threonine.)

使用Phospho-(Ser/Thr) ATM/ATR Substrate Antibody #2851进行ELISA实验:磷酸化和非磷酸化的信噪比。(S*或T*代表磷酸化的丝氨酸或苏氨酸)

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells either untreated or treated with the microtubule destabilizing agents nocodazole or paclitaxol, using Phospho-(Ser) CDKs Substrate Antibody #2324.

对未处理或微管解聚剂噻氨酯哒唑或紫杉醇处理的HeLa细胞抽提液,使用Phospho-(Ser) CDKs Substrate Antibody #2324进行Western blot分析。

Western Blotting

Western Blotting

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-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb.

Western blot分析血清饥饿处理的NIH/3T3细胞抽提物,在体外用AKT激酶磷酸化,或者使用PDGF或者FBS/Calyculin A培养处理,使用的抗体为Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb兔单抗。

IP

IP

Immunoprecipitation of extracts from serum-starved NIH/3T3 cells, untreated or treated with FBS/Calyculin A, using Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb, followed by Western blot analysis using the same antibody.

对血清饥饿后未使用或者使用FBS/Calyculin A处理的NIH/3T3细胞进行免疫沉淀,使用的抗体为Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb兔单抗,然后使用同样的抗体进行Western blot分析。

ELISA-Peptide

ELISA-Peptide

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.)

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。)

IHC-P (paraffin)

IHC-P (paraffin)

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

IHC-P (paraffin)

IHC-P (paraffin)

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

Western Blotting

Western Blotting

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)。

ELISA-Peptide

ELISA-Peptide

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.)

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。)

Western Blotting

Western Blotting

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。

ELISA-Peptide

ELISA-Peptide

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.)

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)

Western Blotting

Western Blotting

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)。

ELISA-Peptide

ELISA-Peptide

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)

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma control (left) or lambda phosphatase-treated (right), using Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb.

免疫组化分析石蜡包埋的人乳腺癌对照(左图)或 lambda磷酸酶处理(右图),使用的抗体为Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb兔单抗。

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untreated (-) or UV-treated (+, 50 mJ/cm2, 2 hr), using Phospho-(Ser/Thr) ATM/ATR Substrate (S*/T*QG) (P-S/T2-100) Rabbit mAb. Western blot was imaged using Odyssey® Infrared Imaging System (LI-COR® Biotechnologies).

对未处理(-)或UV处理(+, 50 mJ/cm2, 2小时)的Hela细胞抽提物进行Western blot分析,使用的抗体为Phospho-(Ser/Thr) ATM/ATR Substrate (S*/T*QG) (P-S/T2-100) Rabbit mAb兔单抗。Western blot图像采集使用Odyssey® Infrared Imaging System (LI-COR® Biotechnologies)。

IP

IP

Immunoprecipitation of Hela cells, untreated (-) or UV-treated (+, 50 mJ/cm2, 2 hr), using Phospho-(Ser/Thr) ATM/ATR Substrate (S*/T*QG) (P-S/T2-100) Rabbit mAb (lanes 3 and 4). Lanes 1 and 2 show 10% input. Western blot detection was performed using the same antibody (upper) and p95/NBS1 Antibody #3002 (lower).

对未处理(-)或UV处理(+, 50 mJ/cm2, 2小时)的Hela细胞抽提物进行免疫沉淀分析,使用的抗体为Phospho-(Ser/Thr) ATM/ATR Substrate (S*/T*QG) (P-S/T2-100) Rabbit mAb 兔单抗(列3、4). 列1、2 是10%输入对照. Western blot检测使用相同的抗体 (上图) 和p95/NBS1 Antibody #3002 (下图)。

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat cells, untreated (-) or TPA-treated (200 nM, 30 min; +), using Phospho-(Ser) PKC Substrate (P-S3-101) Rabbit mAb. Western blot was imaged using Odyssey® Infrared Imaging System (LI-COR® Biotechnologies).

对未处理(-)或TPA处理(200 nM, 30 min; +)的Jurkat细胞抽提物进行Western blot分析,使用的抗体为Phospho-(Ser) PKC Substrate (P-S3-101) Rabbit mAb兔单抗。Western blot图像采集使用Odyssey® Infrared Imaging System (LI-COR® Biotechnologies)。

IP

IP

Immunoprecipitation (IP) of extracts from Jurkat cells, untreated (-) or TPA-treated (200 nM, 30 min; +) (lanes 3 and 4), using Phospho-(Ser) PKC Substrate (P-S3-101) Rabbit mAb. 10% input is shown in lanes 1 and 2. Western blot analysis was performed using the same antibody.

对未处理(-)或TPA处理 (200 nM, 30 min; +)(列3、4)的Jurkat细胞抽提物进行免疫沉淀分析,使用的抗体为Phospho-(Ser) PKC Substrate (P-S3-101) Rabbit mAb兔单抗。列1、2为10%的输入对照。

Western Blotting

Western Blotting

Western blot analysis of extracts from various mouse tissues using Phospho-(Ser) PKC Substrate (P-S3-101) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from H1650 cells, untreated or treated with phenformin (5 mM, 1 hr), using Phospho-(Ser/Thr) AMPK Substrate (P-S/T2-102) Rabbit mAb. Western blot was imaged using Odyssey® Infrared Imaging System (LI-COR® Biosciences).

Western Blotting

Western Blotting

Western blot analysis of extracts from various mouse tissues using Phospho-(Ser/Thr) AMPK Substrate (P-S/T2-102) Rabbit mAb.

IP

IP

Immunoprecipitation of extracts from H1650 cells using Phospho-(Ser/Thr) AMPK Substrate (P-S/T2-102) Rabbit mAb. Lane 1 shows 10% input. Western blot analysis was performed using the same antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from COS-7 cells, synchronized in G1/S, G2/M, and G0 phase of the cell cycle, using Phospho-(Ser) CDKs Substrate (P-S2-100) Rabbit mAb.

ELISA-Peptide

ELISA-Peptide

Phospho-(Ser) CDKs Substrate (P-S2-100) Rabbit mAb ELISA assay: Signal-to-noise ratio of phospho- versus nonphospho-peptides. (S* denotes phosphorylated serine.)

Western Blotting

Western Blotting

Western blot analysis of extracts from COS-7 cells, synchronized in G1/S, G2/M, and G0 phase of the cell cycle, using Phospho-(Ser) CDKs Substrate (P-S2-100) Rabbit mAb.

对COS-7细胞,将其同步在G1/S、G2/M和G0期,使用Phospho-(Ser) CDKs Substrate (P-S2-100) Rabbit mAb进行Western blot分析。

Western Blotting

Western Blotting

Western blot analysis of extracts from H1650 cells, untreated or treated with phenformin (5 mM, 1 hr), using Phospho-(Ser/Thr) AMPK Substrate (P-S/T2-102) Rabbit mAb #5759. Western blot was imaged using Odyssey® Infrared Imaging System (LI-COR® Biosciences).

Background

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.

磷酸化丝氨酸/苏氨酸激酶和磷酸酶在生物过程中扮演重要角色。每个磷酸化丝氨酸/苏氨酸激酶磷酸化特异性基序中的丝氨酸和苏氨酸。Akt仅在底物中存在-5、-3位精氨酸时才能磷酸化丝氨酸或苏氨酸(1)。传统型PKC同工酶磷酸化包含丝氨酸和苏氨酸的底物,要求精氨酸或赖氨酸在-3、-2和+2位,并且疏水氨基酸在+1位(2,3)。PKA的共有磷酸化位点是-2、-3位是精氨酸的丝氨酸或苏氨酸(3)。MAPK和CDK丝氨酸/苏氨酸蛋白激酶家族磷酸化脯氨酸残基之前的苏氨酸或丝氨酸(3-5)。CDK底物的共有氨基酸序列是(K/R)(S*)PX(K/R),X代表任意20个氨基酸之一,S*是磷酸化位点(6-8)。ATM和相关激酶ATR磷酸化S*/T*Q基序中的丝氨酸或苏氨酸(9,10)。某些激酶底物特异性的抗体对于研究激酶活性和发现潜在底物具有很高价值。CST开发了能够识别包含Akt、PKC、PKA、MAPK/CDK、CDK或ATM/ATR可以磷酸化的基序中磷酸化丝氨酸和苏氨酸的这些抗体。通过肽段配对ELISA,此试剂盒中每个磷酸化丝氨酸/苏氨酸激酶底物抗体都有特异性的目标激酶底物基序。

  1. Alessi, D.R. et al. (1996) FEBS Lett 399, 333-8.
  2. Nishikawa, K. et al. (1997) J Biol Chem 272, 952-60.
  3. Pearson, R.B. and Kemp, B.E. (1991) Methods Enzymol 200, 62-81.
  4. Songyang, Z. et al. (1996) Mol Cell Biol 16, 6486-93.
  5. Songyang, Z. (1999) Prog Biophys Mol Biol 71, 359-72.
  6. Holmes, J.K. and Solomon, M.J. (1996) J Biol Chem 271, 25240-6.
  7. Kastan, M.B. and Lim, D.S. (2000) Nat Rev Mol Cell Biol 1, 179-86.
  8. Zhao, H. and Piwnica-Worms, H. (2001) Mol Cell Biol 21, 4129-39.

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U.S. Patent No. 5,675,063.

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