AngiotensinProteinKinaseAssay
James Hardwick's angiotensin assay protocolThis specific procedure was developed to assay the activity of the Lck kinase expressed from a retroviral vector in rat fibroblasts. You can obviously use fewer or more cells, depending on your needs. Don't skimp too much on the amount of S. aureus bugs that you use for precipitation or you will risk losses.1. Immunoprecipitate the protein from 3 x 106 cells using 45......阅读全文
Angiotensin－Protein－Kinase－Assay
James Hardwick's angiotensin assay protocolThis specific procedure was developed to assay the activity of the Lck kinase expressed from a retrovir
HISTONE－KINASE－ASSAY
PROTOCOLTo 1.5 mL eppendorf tubes add:200 µg of protein extract (see Western blot protocol for protein sample preps)q.s. to 300 µL with RIPA (with pro
Protein－Kinase－A－at－the－Centrosome
Protein kinase A regulatory subunit RIIalpha (PKA-RIIa) is tightly bound to centrosomal structures during interphase through interaction with the A-ki
Protein－Assay－(Spectrophotometer)
Protein Assay (Spectrophotometer)Use BSA (bovine serum albumin) 1mg/ml stock solution (1ml Eppendorf tubes) for standard curve.Place 0, 2, 5, 10, 15,
LOWRY－PROTEIN－ASSAY
The Lowry procedure is one of the most venerable and widely-used protein assays, being first described in 1951 [Lowry et al., J. Biol. Chem. 193: 265-
Bradford－protein－assay
Bradford protein assayConsiderations for useThe Bradford assay is very fast and uses about the same amount of protein as the Lowry assay. It is fairly
BIURET－PROTEIN－ASSAY
BIURET PROTEIN ASSAYMATERIALSBiuret ReagentBovine serum albumin (BSA)Spectrophotometer and tubesPROCEDUREPrepare standard dilutions of BSA containing
James－Hardwicks－angiotensin－assay－protocol
This specific procedure was developed to assay the activity of the Lck kinase expressed from a retroviral vector in rat fibroblasts. You can obviousl
Biorad－Protein－Assay:－Bradford
Biorad Protein Assay: BradfordStandards: 1 mg/ml BSA stock- dilute 1:10 to get 0.1 mg/ml BSAAdd To get H-2O20 µl 2 µg/ml 780 µl40 µl 4 µg/ml 760 µl60
Bradford－Protein－Concentration－Assay
Bradford Protein Concentration Assayversion 01/07/2001Abbreviations:mcg = microgramsmcL = microlitersBSA = bovine serum albuminO.D. = optical densityd
In－Vitro－Protein－Ubiquitination－Assay
Ubiquitination is one of the most important posttranslational modifications in all eukaryote organisms. Ubiquitin-activating enzyme (E1), ubiquiti
Activation－of－cAMPdependent－protein－kinase,－PKA
G-protein coupled receptors (GPCRs) are one of the largest gene families of signaling proteins. Residing in the plasma membrane with seven transmembra
Ca++/－Calmodulindependent－Protein－Kinase－Activation
The calcium/calmodulin-dependent kinases (CaMKs) are involved in a large number of cellular responses induced by hormones, neurotransmitters and other
Activation－of－Csk－by－cAMPdependent－Protein－Kinase－Inhibits－Signaling
Interaction of T cell receptor with specific antigen in the context of MHC II activates a signal transduction pathway that leads to T cell activation.
Use－of－the－Bradford－Protein－Assay－in－a－Microtiter－Plate－Format
Introduction The Bradford protein assay is a simple procedure for determination of protein concentrations in solutions that depends upon the change in
Bradford法蛋白定量（Bradford－Protein－Assay－）
Bradford Assay is a rapid and accurate method commonly used to determine the total protein concentration of a sample. The assay is based on the observ
Role－of－EGF－Receptor－Transactivation－by－GPCRs－in－Cardiac－Hypertrophy
One of responses to increased blood pressure is cardiac hypertrophy through increased size of ventricular myocardial cells leading to increased thickn
Identification－of－a－Mutant－Kinase/ATP－Analog－Pair2
Cellular Transfection and Immunoprecipitation Before proceeding with the experiments outlined below, all kinase pocket mutants should be characterize
Bioactive－Peptide－Induced－Signaling－Pathway
Many different peptides act as signaling molecules, including the proinflammatory peptide bradykinin, the protease enzyme thrombin, and the blood pres
蛋白质磷酸化
Tyrosine Kinase Assay Using Synthetic Peptides (T. Miller)Small synthetic peptide substrates are especially well suited for applications such as assay
Role－of－arrestins－in－the－activation－and－targeting－of－MAP－kinases
The binding of ?-arrestins to agonist-occupied GPCRs triggers the assembly of a MAP kinase activation complex using ?-arrestin as a scaffold, with sub
Identification－of－a－Mutant－Kinase/ATP－Analog－Pair1
Identification of a Mutant Kinase/ATP Analog PairScott T. Eblen, N. Vinay Kumar, and Michael J. WeberDepartment of Microbiology and Cancer Center, Uni
Angiotensin－II－mediated－activation－of－JNK－Pathway－via－Pyk2－dependent
Ang II binding to AT1-R triggers the activation of Ca2+ signaling and PKC. The signal is then transmitted to the Pyk2 and further to the small G prote
Penicillan－Binding－Protein－Assay青霉素与细胞膜蛋白结合实验
Wash cells with 10 mM Tris pH 8French pressSlow speed spinHigh speed spinResuspend in 10 mM TrisSonicate 2 x 15 sec to remove ß-lacatamasesWash in 10
SAPK/Jun－kinase－assays
Preparation of cell lysate:1. For cells in suspension, grow in DMEM with 10% FCS and then the day before you do the experiment spin the cells down (1
Human－Cytomegalovirus－and－Map－Kinase－Pathways
To replicate in the host cell, viruses commandeer cellular signaling pathways. Cytomegalovirus (CMV) is a DNA virus with that is widespread in the pop
Map－Kinase－Inactivation－of－SMRT－Corepressor
Corepressors are coregulators that interact with transcriptional silencers in a variety of pathways such as cell proliferation, differentiation and ap
Sprouty－regulation－of－tyrosine－kinase－signals
Four different members of the Sprouty protein family block the cellular proliferation and differentiation induced by several different growth factors,
Antibody－Purification－using－Protein－A,－Protein－G,－or－Protein－L－Agarose
实验概要This protocol is designed as a quick purification method for antibodies from mammalian sera, ascites, and cell culture supernatants主要试剂 Protein
Antibody－Purification－using－Protein－A,－Protein－G,－or－Protein－L－Agarose
实验概要This protocol is designed as a quick purification method for antibodies from mammalian sera, ascites, and cell culture supernatants. It should