HISTONEKINASEASSAY
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 protease and phosphatase inhibitors).Primary antibody (amount determined emperically, approx 10x the concentration needed for westerns)Vortex and incubate on ice for 30 min.Meanwhile wash Protein A-Sepharose beads 2x in RIPA as follows:Cut off ends of pipetman tipsSuspend ......阅读全文
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
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－blotting－protocol
实验概要 Western blot detection of histone proteins. 实验步骤 The following protocol refers to the western blot detection of histone proteins derived from p
Histone－blotting－pr...
实验概要The method provides a procedure and tips for detecting histone proteins.The protocol refers to the western blot detection of histone proteins d
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
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
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,
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
TUNEL－assay
PROTOCOL:•Deparaffinize and rehydrate slides:3 x 3´ Xylene3 x 2´ 100% ethanol1 x 2´ 95%, 80%, 70% ethanol (each)1 x 5´ 1x PBS•Microwave antigen retrie
Aspartate－Assay
实验概要The Aspartate Assay Kit provides a simple, convenient assay to measure aspartate in a variety of samples. In the assay, aspartate is converted
Phosphate－Assay
1. Make standards using sodium phosphate at the following uM concentrations: 0, 2, 5, 7, 10, 20, 40, 60, and 80. Use the screw top glass tubes.2. Dry
Pectinase－assay
Pectinases are actually a mixture of enzymes, which, along with others such as cellulase, are widely used in the fruit juice industry where they are w
Protease－assay
In certain fruits, such as pineapples and mangoes, the flesh contains protein-digesting enzymes (proteases). These may play a part in helping to softe
Polygalacturonase－assay
This enzyme is famous for being involved in the development of the GMO tomatoes (more information from the link at the foot of this page). The cells o
Protease－assay
实验概要 In certain fruits, such as pineapples and mangoes, the flesh contains protein-digesting enzymes (proteases). These may play a part in
MTT－Assay
This procedure is for cells in 96 well plates, if larger plates are used then adjust volumes accordingly.1 Make a solution of 5mg/ml MTT dissolved in
Bradford－Assay
Bradford AssayThe bradford dye-binding assay is a colorimetric assay for measuring total protein concentration. It involves the binding of Coomassie B
DGK－Assay
Buffers:- 2X buffer10 ml 0.5 M imidazol, pH 6.60.21 g LiCl1.25 ml 1 M MgCl21.0 ml 0.1 M EGTA, pH 6.6--> Bring volume up to 50 ml with distilled water.
Motility－Assay
DescriptionVarious phenotypic characteristics are requiredfor a cancer cell to successfully complete the metastaticcascade. Among these, acquisition o
Chemotaxis－Assay
PurposeThe purpose of a chemotaxis assay is to determine whether your protein or small molecule of interest has chemotactic activity on a specific cel
Bradford－Assay
The bradford dye-binding assay is a colorimetric assay for measuring total protein concentration. It involves the binding of Coomassie Brilliant blue
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
Assay－of－Phospholipase－A－Activity
Phospholipases of the A type constitute a large family of esterases that catalyze the hydrolysis of the fatty acid ester bonds in phospholipids an
Actin－Capture－Assay
David AmbergDialyze purified GST fusion proteins and actin into PBS + 1mM MgCl2 .Mix 5ug actin into 50ul total volume binding buffer.Mix 5ug GST-fusio
Crystal－Violet－Assay
This is a simple assay useful for obtaining quantitative information about the relative density of cells adhering to multi-well cluster dishes. The dy
Pheromone－Halo－Assay
-Use sterile technique and sterile solutions throughout this method.-1. Grow a starter culture at 30 C with shaking (250 rpm) until it reaches saturat
Assay－for－the－Micrococcal－Nuclease
Method: Essentially that described by Heins et al. (1966) based upon the release of acid soluble oligonucleotides following nuclease digestion of DNA.
Migration－Assay－Protocol
Materials to be prepared beforehand:1) FBS free medium2) 10% FBS medium3) Cell migration filter insert ( Transwell®, 12mm Diameter, 12 μm Pore Size.)P
In－vitro－Sphingomyelinase－Assay
Reagents:Lysis buffer25 mM Tris-HCl, pH 7.45 mM EDTA1 mM ATP20 µg/ml CLAP1 mM PMSFBuffer A10 mM MgCl20.2 M Tris-HCl, pH 7.40.2 % Triton X-100Buffer B0