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(PI 3-Kinase) Activity Assay - http://www.upstate.com/misc/protocol_detail.q.prot.e.assay.a.name.e.Phosphatidyl_Inositol-3_Kinase_Assay_%28PI3%20Kinase%20Assay%29

Category: Signalling Signal Transduction Protocols

Protocol describing (PI 3-Kinase) activity assay. Includes Preparation of Phosphatidylinositol (PI). - [Read (PI 3-Kinase) Activity Assay]

Activated MAP Kinase Protocols Stratagene PDF - http://www.stratagene.com/manuals/206110.pdf

Category: Signalling Signal Transduction Protocols: MAPK Protocols

MAPK Protocols. Resolving Phosphorylated MAPK Proteins by SDSâ€“PAGE, Filter Binding Assay for activated MAPK Î³-32P incorporation of MAPK. Stratagene - [Read Activated MAP Kinase Protocols Stratagene PDF]

Activation of p70s6k and MAP Kinases: IP and Kinase Activity - http://www.whitelabs.org/Lab%20Protocols/kinase%20and%20phosphatase%20assays/ACTIVATION%20OF%20p70s6k%20and%20MAP%20KINASES.htm

Category: Signalling Signal Transduction Protocols: MAPK Protocols

Protocol describes the activation of MAP Kinase activity. Protocol contains information on: Buffers: ST, STE, Lysis Buffer Buffer A, Buffer B and 10x Kinase Buffer. - [Read Activation of p70s6k and MAP Kinases: IP and Kinase Activity]

Akt/PKB Kinase Assay - http://www.whitelabs.org/Lab%20Protocols/kinase%20and%20phosphatase%20assays/Akt%20PKB%20kinase%20assay.htm

Category: Signalling Signal Transduction Protocols

Protocol for Akt/PKB kinase assay. Protocol includes: Akt lysis buffer; Kinase Buffer; 1.5x Akt kinase reaction buffer; Start solution; Lipofectamine PLUS; CDNA synthesis (Roche #1483?88). - [Read Akt/PKB Kinase Assay]

Analysis of Membrane Trafficking and Intracellular Signaling in Self-Generated Iodixanol Gradients - http://www.axis-shield.com/densityhome/optiprep/S25.pdf

Category: Protein Protocols: Protein Trafficking

Protocol is based on methods for the resolution of GLUT4 containing vesicles and the identification of phosphoinositide kinase containing vesicles in 3T3-L1 adipocytes. They may have a wider application to any low-medium density membranes. Protocol incorporates the strategy of using a low density microsome fraction as the gradient input, commonly used in GLUT 4 studies that may have a wider application to other investigations. - [Read Analysis of Membrane Trafficking and Intracellular Signaling in Self-Generated Iodixanol Gradients]

Analysis of protein phosphorylation: methods and strategies PDF - http://www.blackwell-synergy.com/doi/pdf/10.1111/j.1365-313X.2005.02613.x

Category: Protein Protocols: Protein Phosphorylation Protocols

Paper describing methods for monitoring kinase activity, investigating kinaseâ€“substrate specificity, examining phosphorylation in planta and the determination of phosphorylation sites in a protein. In addition, strategic considerations for experimental design and variables will be discussed. Scott C. Peck, the Plant Journal. - [Read Analysis of protein phosphorylation: methods and strategies PDF]

Angiotensin assay protocol - http://pingu.salk.edu/~sefton/Hyper_protocols/angio.html

Category: Signalling Signal Transduction Protocols: Protein Kinase Protocols

Angiotensin assay protocol. Bart's Cookbook. Assay activity of Lck kinase expressed in a retroviral vector in rat fibroblasts. - [Read Angiotensin assay protocol]

Assay for Immunoprecipitated Phosphoinositide 3-Kinase (PI 3 Kinase) Activity (in vitro) - http://www.upstate.com/misc/protocols.q.prot.e.assay/Assay+for+Immunoprecipitated+Phosphoinositide+3+Kinase++PI+3+Kinase++Activity+%3Ci%3E+in+vitro+%3C%2Fi%3E

Category: Signalling Signal Transduction Protocols: Protein Kinase Protocols

Immunoprecipitation assay for Phosphoinositide 3-Kinase (PI 3 Kinase) Activity (in vitro) Protocol. Upstate. - [Read Assay for Immunoprecipitated Phosphoinositide 3-Kinase (PI 3 Kinase) Activity (in vitro)]

CAT Assay Protocol - http://kinase.uhnres.utoronto.ca/methods/CATassay.html

Category: Reporter Gene Assays: Cat Assays Protocols

Protocol for CAT assay. - [Read CAT Assay Protocol]

Detailed NF-kB EMSA Protocol - http://www.celldeath.de/apometh/emsa.html

Category: Cell Biology and Cell Culture: Apoptosis Protocols: NF-kB EMSA Protocols

Detailed NF-kB EMSA Protocol. Includes Nuclear extract preparation, Labeling of the oligonucleotide probe: T4 Polynucleotide Kinase Reaction, Supershift assay to identify NF-kB, and recipes for EMSA buffers. Celldeath.de - [Read Detailed NF-kB EMSA Protocol]

FAK Autophosphorylation Assay - http://www.bio.com/protocolstools/protocol.jhtml?id=p1984

Category: Signalling Signal Transduction Protocols

This kinase assay is meant to determine whether an agonist or event can influence the autophosphorylation of FAK. The addition of 1 Î¼l of polyGT to the kinase reaction mix will determine the activity of the enzyme against a substrate. Includes information on: Harvest, Immunoprecipitation, Kinase Reaction and Antibody Detection of FAK. - [Read FAK Autophosphorylation Assay]

Histone H1 Kinase Activity Assay - http://www.bio.com/protocolstools/protocol.jhtml?id=p910

Category: Signalling Signal Transduction Protocols

This protocol describes assaying kinase activity of a putative kinase using Histone H1 as the substrate. Histone H1 is the canonical kinase substrate in this type of assay. Phosphorylation of Histone H1 is assessed by SDS-Polyacrylamide gel electrophoresis followed by autoradiography. Includes information: H1 Kinase Assay on Individual Xenopus Oocytes; H1 Kinase Assay on Xenopus Egg Extract Samples; H1 Kinase Assay on Tissue Culture Cells; Helpful protocol hints. - [Read Histone H1 Kinase Activity Assay]

Immunoprecipitation / Kinase Assay - http://www.upstate.com/misc/protocols.q.prot.e.ipkinaseassay/Immunoprecipitation+%2F+Kinase+Assay

Category: Signalling Signal Transduction Protocols: Protein Kinase Protocols

Immunoprecipitation / Kinase Assay Protocol. Upstate. Protocol applicable only to kinases whose activity is not altered by cell lysis or immunoprecipitation procedures, and do not require soluble cofactors for activity. - [Read Immunoprecipitation / Kinase Assay]

Immunoprecipitation of Endogenous MEKK1 and Kinase Assay Protocol - http://www.natureprotocols.com/2007/01/22/immunoprecipitation_of_endogen.php

Category: Immunology: Immunoprecipitation Protocols: Protein Immunoprecipitation Protocols

Protocol for immunoprecipitation of endogenous MEKK1 and kinase assay. Includes: Preparation of lysates; Immunoprecipitation; Kinase assay. - [Read Immunoprecipitation of Endogenous MEKK1 and Kinase Assay Protocol]

In Vitro MAP Kinase Assay - http://www.whitelabs.org/Lab%20Protocols/kinase%20and%20phosphatase%20assays/In%20Vitro%20MAP%20kinase%20assay%20%209.htm

Category: Signalling Signal Transduction Protocols: MAPK Protocols

Protocol describes an in vitro assay to detect MAP kinase activity using PC12 cells. - [Read In Vitro MAP Kinase Assay]

In Vitro Transcription With Yeast Nuclear Extract - http://www.fhcrc.org/science/labs/hahn/methods/biochem_meth/polii_trans.html

Category: Cell Biology and Cell Culture: In Vitro Reconstitution Protocols

Protocol for in vitro transcription with yeast nuclear extract. Includes recipes for: 5x Acetate transcription buffer; Creatine phospho kinase; Phospho creatine; Stop mix; HA + 0.1 M potassium acetate; 5x glutamate transcription buffer (5 ml). Includes also protocol for Primer Extension Assay. - [Read In Vitro Transcription With Yeast Nuclear Extract]

In Vivo Functional Real-Time Imaging of Transplanted Islets Using Positron Emission Tomography (PET) - http://www.natureprotocols.com/2006/12/21/seeing_is_believing_in_vivo_fu.php

Category: Microscopy Protocols: In Vivo Imaging Protocols

Positron emission tomography (PET) is a established quantitative and noninvasive imaging modality. With the PET reporter gene (PRG)/PET reporter probe (PRP) system, based on a mutant form of herpes simplex virus 1 thymidine kinase (HSV1-sr39tk), the PET signal is directly proportional to the enzymatic activity of sr39TK9-14. In this protocol, we describe in detail a method for reporter gene labeling of islets and quantitative scanning using a reporter probe. - [Read In Vivo Functional Real-Time Imaging of Transplanted Islets Using Positron Emission Tomography (PET)]

In-Gel Kinase Assay - http://www.bio.com/protocolstools/protocol.jhtml?id=p911

Category: Signalling Signal Transduction Protocols

Protocol describes how to assay for kinase activity within a polyacrylamide gel, rather than in solution. The advantages to an in-gel assay are that an apparent molecular weight can be assigned to the kinase activity and that multiple kinase activities can be distinguished. Includes protocol hints. - [Read In-Gel Kinase Assay]

In-Vitro Kinase Assay for Jnk-1/IRS Proteins - http://www.whitelabs.org/Lab%20Protocols/kinase%20and%20phosphatase%20assays/In_vitro%20kinase%20assay%20for%20JNK-1%20IRS%20proteins.htm

Category: Signalling Signal Transduction Protocols: MAPK Protocols

Protocol describing an in vitro assay on the detection of Kinase activity for Jnk-1/IRS proteins. - [Read In-Vitro Kinase Assay for Jnk-1/IRS Proteins]

JNK Kinase Assay - http://www.whitelabs.org/Lab%20Protocols/kinase%20and%20phosphatase%20assays/JNK%20Kinase%20Assay%20LRui.htm

Category: Signalling Signal Transduction Protocols: MAPK Protocols

Protocol describing an JNK Kinase Assay. Protocol includes information on reaction buffer. - [Read JNK Kinase Assay]

Kinase Assay Using In Vitro Translated Xenopus Mos Kinase - http://www.bio.com/protocolstools/protocol.jhtml?id=p912

Category: Signalling Signal Transduction Protocols

Protocol describes how tagged Mos kinase is translated in vitro, immunopurified, and used in a kinase assay. Protocol includes: Translation of Xenopus Mos Kinase; Antibody to Antigen Binding; Protein A Sepharose to Antibody Binding; Kinase Reactions on Immunoprecipitated Material; Polyacrylamide Gel Analysis of Immunoprecipitates. Includes protocol hints. - [Read Kinase Assay Using In Vitro Translated Xenopus Mos Kinase]

Kinase Assays - http://www.natureprotocols.com/2007/01/29/kinase_assays.php

Category: Signalling Signal Transduction Protocols: MAPK Protocols

Nature protocol entailing a kinase assay. - [Read Kinase Assays]

Lambda gt22a Screening Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/L1.html

Category: Viral Manipulation Protocols: Phage Lamda Manipulation Protocols

Protocol for lambda gt22a screening. Includes: T-TYN Media + Mg+2; T-TYN Plates; T-TYN Top Agarose; Phage Dilution Buffer (PDB); 10X Kinase Buffer (-DTT); Calf Thymus DNA. - [Read Lambda gt22a Screening Protocol]

Mitogen Activated Protein Kinase Guide - http://www.promega.com/guides/sigtrans_guide/strg_chap01.pdf

Category: Signalling Signal Transduction Protocols: MAPK Protocols

Mitogen Activated Protein Kinase Guide. MEK, MEKK, MAPK, MAPKK, antibodies and kits Promega. - [Read Mitogen Activated Protein Kinase Guide]

Phosphoamino acid analysis of non-TCA precipitable proteins - http://kinase.uhnres.utoronto.ca/methods/PAA.html

Category: Protein Protocols: Protein Quantitation Concentration Protocols

Phosphoamino acid analysis of non-TCA precipitable proteins and Alkali treatment of gels protocols. Woodgett Lab. - [Read Phosphoamino acid analysis of non-TCA precipitable proteins]

Articles (1-2 of 2)

In Vitro Translated Xenopus Mos Kinase Assay Protocol

In Vitro Translated Xenopus Mos Kinase Assay Protocol. In response to progesterone, immature Xenopus oocytes mature to eggs that can be fertilized. The Mos protein kinase is essential for oocyte maturation, most likely due to its ability to activate the MAP kinase cascade. This MAP kinase cascade eventually leads to the activation of Cdc2/cyclin B and entry into M phase. In this protocol, tagged Mos kinase is translated in vitro, immunopurified, and used in a kinase assay.

Histone H1 Kinase Activity Assay

Histone H1 Kinase Activity Assay Protocol. This protocol describes assaying kinase activity of a putative kinase using Histone H1 as the substrate. Histone H1 is the canonical kinase substrate in this type of assay. Phosphorylation of Histone H1 is assessed by SDS-Polyacrylamide gel electrophoresis followed by autoradiography.

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