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
BIURET－PROTEIN－ASSAY
BIURET PROTEIN ASSAYMATERIALSBiuret ReagentBovine serum albumin (BSA)Spectrophotometer and tubesPROCEDUREPrepare standard dilutions of BSA containing
DNA－methyltransferase－Assay
Methylated CpG island Amplification Protocol written by Minoru Toyota2. Materials2.1. MCARestriction enzymes SmaI, XmaIT4 DNA ligaseTaq DNA polymerase
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
Adhesion－Assay－Protocol
Materials to be prepared beforehand:1) Washing Buffer--0.1% BSA in medium (DMEM or RPMI)2) Blocking Buffer--0.5% BSA in medium (DMEM or RPMI)3) Lamini
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
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
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,
Cell－Viability－Assay
Dye exclusiona cell suspension is mixed with trypan blue and examined by low-power microscopyMaterialscellsPBSM3hemocytometer0.4 % trypan blue in PBSm
cell－proliferation－assay
cell proliferation assaybefore start:thaw cells from liquid nitrogen, grow in 75cc flask (T75) in Fischer's medium MM (maintenance medium) until c
ELISA－Inhibition－Assay
ELISA Inhibition AssaySensitize a 96-well microtiter plate with purified antigen.Prepare a solution of the purified antigen of interest in phosphate b
Wound－healing－assay
The wound healing assay allows the researcher to study cell migration and cell interactions. In some cases also single cell migration can be analyzed.
Leaf－GUS－Assay
实验概要a protocol for Leaf GUS Assay This protocol is for small samples (usually single leaf from 21DAI plants), scale up for larger samplesAs there are
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
Glycolipid－Binding－Assay
Glycolipid Binding AssaySource: Contributed by Pingsunjim, Paller’s LabAbstract: This protocol can be used for the detection of glycolipids binding to
Soft－Agar－Assay
Soft Agar AssayMake 0.6% media-agar mix for the bottom layer. To make 0.6% agar mix the following components (this makes 200 ml):2X DME 100 mlI
Leaf－GUS－Assay
一、实验试剂 GUS Buffer (500 ml) 2.0478 g Na2HPO4 1.2688 g NaH2PO4 (=50 mM NaPi pH7.0) 10 ml 0.5 M EDTA (=10 mM) 0.5 g Triton X-100 0.5 g N-L
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
Needle－Assay－for－Chemotaxis
Devreotes Lab, John Hopkins Medical Institutions http://www.hopkinsmedicine.org/cellbio/devreotes/needle.htmEquipment and chemicalsZeiss inverted micr
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
Regulation－of－MAP－Kinase－Pathways－Through－Dual－Specificity－Phosphatases
Mitogen-activated protein (MAP) kinases are important players in signal transduction pathways activated by a range of stimuli and mediate a number of
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
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
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
Control－of－skeletal－myogenesis－by－HDAC－calcium/calmodulindependent－kinase
The differentiation of muscle cells is transcriptionally regulated, in part by the myocyte enhancer factor-2, MEF2. During myogenesis MEF2 binds to My
Hanging－drop－aggregation－assay
DescriptionThis assay is used for the aggregation property of cancer cells. It is a very critical parameter for measurement of cell metastasis. Factor
Polyphenoloxidase－(catechol－oxidases)－assay
Browning of the cut surface of some fruits and vegetables is due the presence of a group of enzymes called polyphenoloxidases. These enzymes are relea
MINICHROMOSOME－MICROTUBULE－BINDING－ASSAY
Determine the OD600 and correlate the cell density from the chart. Set up four 100mL YPD cultures at the following densities: 0.7x105, 1x105, and 3.0x
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
Nucleotide－Binding/Hydrolysis－Assay
MaterialsNucleotide mixMotor (50 - 100 µM; purity > 95%)0.5 M Tris-OAc, pH 7.510 mM EGTA10 mM MgCl2DDWSephadex G-50 Medium column (0.8 cm in x 20 cm)C
GST－Activity－Fluorometric－Assay
实验概要The experiment provides a simple, fluorescence-based in vitro assay for detecting the GST activity using a fluorescence plate reader. The assay