MinichromosomeMicrotubuleBindingAssay微染色體－微管結合實驗1
Koshland Lab,Carnegie Institute http://www.ciwemb.edu/labs/koshland/Protocols/MICROTUBULE/mmb.htmlDetermine the OD600 and correlate the cell density from the chart. Set up four 100mL YPD cultures at the following densities: 0.7x105, 1x105, and 3.0x105 cells/mLGROWTH OF CELLSGrow 100mL of cells to OD600=0.7-0.8 at 23oC.Add 5-10ul of BME, 15 minutes before spin.Harvest cells in two 50mL conical tubes, sp......閱讀全文
Glycolipid－Binding－Assay
Glycolipid Binding AssaySource:?Contributed by Pingsunjim, Paller’s LabAbstract:?This protocol can be used for the detection of glycolipids binding to
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
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
MINICHROMOSOME－MICROTUBULE－BINDING－ASSAY2
HYBRIDIZATION.Prehybridize blot at 65oC for ~3h in Church buffer containing 0.5mg/ml denature salmon sperm DNA (usually 14ml Church buffer plus 0.7ml
MinichromosomeMicrotubule－Binding－Assay－微染色體－微管結合實驗2
YWB per 10ml5mL 2M Sorbitol (if NZ arrested, add 40uL 1.5mg/mL0.336mL 1M K2HPO4?N2 to 4mL YWB)0.064mL 1M KH2PO44.6mL dH2OYWB, glycerol, PMSF5mL 2M Sor
MinichromosomeMicrotubule－Binding－Assay－微染色體－微管結合實驗1
Koshland Lab,Carnegie Institute?http://www.ciwemb.edu/labs/koshland/Protocols/MICROTUBULE/mmb.htmlDetermine the OD600?and correlate the cell density f
Penicillan－Binding－Protein－Assay青霉素與細胞膜蛋白結合實驗
Wash cells with 10 mM Tris pH 8French pressSlow speed spinHigh speed spinResuspend in 10 mM TrisSonicate 2 x 15 sec to remove ?-lacatamasesWash in 10
Microtubule－Binding－Assays
MaterialsSiliconized ultracentrifuge microfuge tubesGTP-depleted microtubules6X SDS loading dye1X SDS loading dyeCoomassie Brilliant Blue R 250 (0.8%
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
Protease－assay
實驗概要? ? ? ? In certain fruits, such as pineapples and mangoes, the flesh contains protein-digesting enzymes (proteases). These may play a part in
Aspartate－Assay
實驗概要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
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.
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
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
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
Bradford－Assay
Bradford AssayThe bradford dye-binding assay is a colorimetric assay for measuring total protein concentration. It involves the binding of Coomassie B
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
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
Motility－Assay
DescriptionVarious phenotypic characteristics are requiredfor a cancer cell to successfully complete the metastaticcascade. Among these, acquisition o
Bradford－Assay
The bradford dye-binding assay is a colorimetric assay for measuring total protein concentration. It involves the binding of Coomassie Brilliant blue
IASYSbinding－cuvette－and－getting－KD
Immobilization of ligands on cuvette surfaces and measure the interactions of ligand which is immobilized on the cuvette and the ligate which is a
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-
Glucosamine－Rapid－Assay
Glucosamine Rapid AssayMRTHOD:Place sample (containing 0.5 - 10 μg GlcN) in a Pyrex screw capped tube.Add HCl to a final concentration of 2N and a fin
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
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.
Noble－Agar－Assay
DescriptionCancer cells do not show anchorage and contact inhibition of growth. To assess the anchorage and contact independent growth of cells, noble
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
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