Revision 1

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Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Product Includes Product # Quantity Mol. Wt Isotype/Source
Phospho-AMPK Substrate Motif [LXRXX(pS/pT) MultiMab® Rabbit Monoclonal Antibody mix575920 µlRabbit
Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit Monoclonal Antibody961420 µlRabbit IgG
Phospho-PKA Substrate (RRXS*/T*) (100G7E) Rabbit Monoclonal Antibody962420 µlRabbit IgG
Phospho-ATM/ATR Substrate Motif [(pS/pT) QG] MultiMab® Rabbit Monoclonal Antibody mix696620 µlRabbit IgG
Phospho-PKC Substrate Motif [(R/K)XpSX(R/K)] MultiMab® Rabbit Monoclonal Antibody mix696720 µlRabbit IgG
Phospho-CDK Substrate Motif [(K/H)pSP] MultiMab® Rabbit Monoclonal Antibody mix947720 µlRabbit IgG
Anti-rabbit IgG, HRP-linked Antibody7074100 µlGoat

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The Phospho-(Ser/Thr) Kinase Substrate Antibody Sampler Kit provides an economical means to investigate the downstream activity of select serine/threonine kinases. The kit contains enough primary antibody to perform two western blot experiments with each antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

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). AMPK phosphorylates consensus motif (L/M)XRXX(S/T)XXXL (6). Antibodies recognizing the LXRXX(S/T) motif are very useful in the identification of AMPK substrates. 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 (4-6). ATM and the related kinase ATR phosphorylate serine or threonine in an S*/T*Q motif (7,8).
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.

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Western blot analysis of extracts from various mouse tissues using Phospho-AMPK Substrate Motif [LXRXX(pS/pT) MultiMab® Rabbit mAb mix
undefined Image 1: Phospho-(Ser/Thr) Kinase Substrate Antibody Sampler Kit
Western blot analysis of extracts from HeLa cells, untreated (-) or UV-treated (+, 50 mJ/cm2, 2 hr), usingPhospho-ATM/ATR Substrate Motif [(pS/pT) QG] MultiMab® Rabbit mAb mix. Western blot was imaged using Odyssey® Infrared Imaging System (LI-COR® Biotechnologies).
undefined Image 2: Phospho-(Ser/Thr) Kinase Substrate Antibody Sampler Kit
Western blot analysis of extracts from Jurkat cells, untreated (-) or TPA-treated (200 nM, 30 min; +), using Phospho-PKC Substrate Motif [(R/K)XpSX(R/K)] MultiMab® Rabbit mAb mix. Western blot was imaged using Odyssey® Infrared Imaging System (LI-COR® Biotechnologies).
undefined Image 3: Phospho-(Ser/Thr) Kinase Substrate Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Flow cytometric analysis of fixed and permeabilized Jurkat cells treated post-fixation with λ-phosphatase (blue) or untreated (green), using Phospho-PKC Substrate Motif [(R/K)XpSX(R/K)] MultiMab® Rabbit mAb mix (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
undefined Image 4: Phospho-(Ser/Thr) Kinase Substrate Antibody Sampler Kit
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.
undefined Image 5: Phospho-(Ser/Thr) Kinase Substrate Antibody Sampler Kit
Western blot analysis of extracts from COS-7 cells, synchronized in G1/S, G2/M, and G0 phase of the cell cycle, usingPhospho-CDK Substrate Motif [(K/H)pSP] MultiMab® Rabbit mAb mix.
undefined Image 6: Phospho-(Ser/Thr) Kinase Substrate Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
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.
undefined Image 7: Phospho-(Ser/Thr) Kinase Substrate Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded lung carcinoma, using Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit Monoclonal Antibody.
undefined Image 8: Phospho-(Ser/Thr) Kinase Substrate Antibody Sampler Kit
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).
undefined Image 9: Phospho-(Ser/Thr) Kinase Substrate Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Western blot analysis of extracts from H1650 cells, untreated or treated with phenformin (5 mM, 1 hr), usingPhospho-AMPK Substrate Motif [LXRXX(pS/pT) MultiMab® Rabbit mAb mix. Western blot was imaged using Odyssey® Infrared Imaging System (LI-COR® Biosciences).
undefined Image 10: Phospho-(Ser/Thr) Kinase Substrate Antibody Sampler Kit
Immunoprecipitation of Hela cells, untreated (-) or UV-treated (+, 50 mJ/cm2, 2 hr), usingPhospho-ATM/ATR Substrate Motif [(pS/pT) QG] MultiMab® Rabbit mAb mix (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).
undefined Image 11: Phospho-(Ser/Thr) Kinase Substrate Antibody Sampler Kit
Western blot analysis of extracts from various mouse tissues using Phospho-PKC Substrate Motif [(R/K)XpSX(R/K)] MultiMab® Rabbit mAb mix.
undefined Image 12: Phospho-(Ser/Thr) Kinase Substrate Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Flow cytometric analysis of fixed and permeabilized Jurkat cells, untreated (blue) or treated with TPA (200 nM, 30 min; green), using Phospho-PKC Substrate Motif [(R/K)XpSX(R/K)] MultiMab® Rabbit mAb mix (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
undefined Image 13: Phospho-(Ser/Thr) Kinase Substrate Antibody Sampler Kit
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.
undefined Image 14: Phospho-(Ser/Thr) Kinase Substrate Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded renal cell carcinoma, using Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb.
undefined Image 15: Phospho-(Ser/Thr) Kinase Substrate Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Immunoprecipitation of extracts from H1650 cells using Phospho-AMPK Substrate Motif [LXRXX(pS/pT) MultiMab® Rabbit mAb mix. Lane 1 shows 10% input. Western blot analysis was performed using the same antibody.
undefined Image 16: Phospho-(Ser/Thr) Kinase Substrate Antibody Sampler Kit
Immunoprecipitation (IP) of extracts from Jurkat cells, untreated (-) or TPA-treated (200 nM, 30 min; +) (lanes 3 and 4), using Phospho-PKC Substrate Motif [(R/K)XpSX(R/K)] MultiMab® Rabbit mAb mix. 10% input is shown in lanes 1 and 2. Western blot analysis was performed using the same antibody.
undefined Image 17: Phospho-(Ser/Thr) Kinase Substrate Antibody Sampler Kit
Phospho-CDK Substrate Motif [(K/H)pSP] MultiMab® Rabbit mAb mix ELISA assay: Signal-to-noise ratio of phospho- versus nonphospho-peptides. (S* denotes phosphorylated serine.)
undefined Image 18: Phospho-(Ser/Thr) Kinase Substrate Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human breast carcinoma control (left) or lambda phosphatase-treated (right), using Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb.
undefined Image 19: Phospho-(Ser/Thr) Kinase Substrate Antibody Sampler Kit
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.)
undefined Image 20: Phospho-(Ser/Thr) Kinase Substrate Antibody Sampler Kit
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.)
undefined Image 21: Phospho-(Ser/Thr) Kinase Substrate Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human breast carcinoma control (left) or lambda phosphatase-treated (right), using Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb.
undefined Image 22: Phospho-(Ser/Thr) Kinase Substrate Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded lung carcinoma, using Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb.
undefined Image 23: Phospho-(Ser/Thr) Kinase Substrate Antibody Sampler Kit
Immunohistochemical analysis of paraffin-embedded renal cell carcinoma, using Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb.
undefined Image 24: Phospho-(Ser/Thr) Kinase Substrate Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines untreated (-) or treated (+) with hEGF (100ng/ml, 4 hours), PDGF (100ng/ml, 5 minutes), calf intestinal phosphatase (CIP)/Lambda Phosphatase, or UV (50mJ/cm2, 1 hour recovery) using Phospho-Akt Substrate (RXXS*/T*) (110B7E) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174.
undefined Image 25: Phospho-(Ser/Thr) Kinase Substrate Antibody Sampler Kit
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.