SAPK／Junkinaseassays
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 K for 5 min) and obtain a concentration of cells at 2 - 10 x 106 cells per sample, resuspended in DMEM, 2 % FCS. This starves the cells or decreases the background. Then incubate at 37oC, overnight or 16 hrs. For adherent cells, simply change the medium to 2% FCS the ni......閱讀全文
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
Agglutination－Assays
Agglutination Assays REFERENCE:?Lanyi, B., and T. Bergan. Methods in? Microbiology, Vol 10: 93-168.? BACTERIAL AGGLUTINATION:? Bacterial agglutinat
Carbohydrate－Assays
Carbohydrate Assays REFERENCE:?Wright and Rebers, Anal. Biochem. 49: 307-319, 1972. OBJECTIVE:?To determine the relative amounts of LPS carbohydrat
Cellulase－assays
Cellulase enzymes show activity during the ripening of some fruits, where their effects on cell walls results in softening of the fruit. In cases of p
Cellulase－assays
實驗概要? ? ? ? Cellulase enzymes show activity during the ripening of some fruits, where their effects on cell walls results in softening of the frui
Protein－Kinase－A－at－the－Centrosome
Protein kinase A regulatory subunit RIIalpha (PKA-RIIa) is tightly bound to centrosomal structures during interphase through
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
Cytotoxicity－Assays－Protocol
Cytotoxicity Assays ProtocolCell-mediated cytotoxicity was determined by using a standard microcytotoxicity assay. Briefly, target cells were pelleted
Matrigel－invasion－assays
OverviewMatirgel is considered as basement membrane and generated from EHS sarcoma. Matrigel contains not only basement membrane components (collagens
Microtubule－Binding－Assays
MaterialsSiliconized ultracentrifuge microfuge tubesGTP-depleted microtubules6X SDS loading dye1X SDS loading dyeCoomassie Brilliant Blue R 250 (0.8%
Angiotensin－Protein－Kinase－Assay
James Hardwick's angiotensin assay protocol This specific procedure was developed to assay the activity of the Lck kinase expressed from a retroviral
PLAQUE－ASSAYS－FOR－ADENOVIRUS－TITRATION
-Set up 60 mm dishes of P11 cells to be 100 confluent at time of infection.?-Remove medium from dishes, add 0.2 to 0.5 ml virus and adsorb for 30 – 60
Coimmunoprecipitation－assays
co-IP assays can be performed between endogenous proteins or transiently or stably expressed exogenous - usually tagged - proteins. The advantage to u
cAMP分析－cAMP－Assays
cAMP AssaysGouzel Karimova and Daniel LadantUnite Postulante de Biochimie des Interactions Macromoleculaires, Departement de Biologie Structurale et C
DNA－Fragmentation－Assays－for－Apoptosis
Protocol I:?Triton X-100 Lysis BufferIn 96 flat-wells plate, incubate 4x10 6 target cells (40 wells of 105 per well) with desired concentration of eff
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
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
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,
Microtubule－(MT)/Organelle－Motility－Assays
Rapidly thaw and immediately place on ice one aliquot each of axonemes, Golgi or ER membranes, 45 uM tubulin, rat liver cytosol, and 20x energy regene
Nuclear－RunOn－Transcription－Assays
Nuclear “run-on” (or “run-off”) transcription assays have been used to obtain quantitative information about the relative rates of transcription o
Phosphatidylinositol－4Kinase－and－Phosphatidylinositol－4Phosphate－5K...
Phosphatidylinositol 4-Kinase and Phosphatidylinositol 4-Phosphate 5-Kinase Assays Inositol lipid kinases are perhaps the easiest and most straightf
Fragment－Complementation－and－Coimmunoprecipitation－Assays－for－...
Plant disease resistance (R) proteins confer protection against specific pathogens or pathogen isolates. R proteins function by recognizing patho
Preparation－of－Luciferin－for－In－Vitro－and－In－Vivo－Bioluminescent－Assays
Preparation of Luciferin forIn Vitro Bioluminescent AssaysMaterials? D-Luciferin Firefly, potassium salt, 1.0 g /vial(Caliper Life Sciences Part Numbe
ATPase－Assays－with－32PATP
MaterialsPurified Motor Protein, 20-80 μMNucleotide Mix =50 mM Mg·ATP?gamma-32P-ATP to give 5 000 - 10 000 cpm/nmol?10 mM HEPES, pH 7.2?1 mM EGTA?1 mM
Preparation－of－Luciferin－for－In－Vitro－and－In－Vivo－Bioluminescent－Assays
實驗概要Reagent ?for immunoassay, ligand binding assay and ligand receptor assay in ?which luciferin is covalently bonded to a molecule having biological
Preparation－of－Luciferin－for－In－Vitro－and－In－Vivo－Bioluminescent－Assays
實驗概要Reagent ?for immunoassay, ligand binding assay and ligand receptor assay in ?which luciferin is covalently bonded to a molecule having biological
蛋白質磷酸化
Tyrosine Kinase Assay Using Synthetic Peptides?(T. Miller)Small synthetic peptide substrates are especially well suited for applications such as assay
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
Flow－Cell－Assays－with－Microtubules:－Motility/Dynamics－in－Fluorescence
Flow cell assays are very useful for studying microtubule motility, microtubule dynamics, kinetochore-microtubule interactions and action of severing
Steady－State－ATPase－Assays－Coupled－Enzyme－System
MaterialsTubulin?(>5 mg/mL)100 mM Mg·GTP4 mM Taxol in DMSOPM =100 mM PIPES pH 6.82 mM EGTA1 mM Mg2SO4Motor protein (>95% purity; 15-20 μM)Cuvettes (20