化学生物学实验技术

Amicon Stirred Ultrafiltration Cells (Models 8050 8400) Description - Membranes - Assembly - Operation - Limits - Sterilization - Maintenance - Troubleshooting DescriptionFor protein concentration gas ...

James Hardwick's 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 obviously use f ...

Biosynthetic labelingHow long should cells be labeled? The ideal length of time to label cells depends on the protein of interest and the label that you are using. If you want to label an unstable pro ...

Liposome Swelling AssayREFERENCES: Nikaido and Roseenberg (1983) J.Bact 153: 241-252; Yoshimura et al. (1983) J.Bact 258: 2308-2314.METHOD:6.2 µmoles of lipid (ideally egg phophatidylcholine or dipalm ...

Assay of superoxide dismutase activity by combining electrophoresis and densitometryAbstract. A modified technique was developed to assay superoxide dismutase (SOD) activity by combining polyacrylamid ...

Permeabilization of gram-negative bacteria with KPi/hexane for the release of L-asparaginaseSource: Hikmet Geckil's LabAbstract: This is a fast efficient and reproducible recovery procedure for peripl ...

Phosphoamino acid analysis:Mark Kamps's method 1. Label your protein with 32Pi. Then subject the protein to SDS polyacrylamide gel electrophoresis and transfer your gel-fractionated protein to Immobil ...

Preparing a Selenomethionyl Proteinby Sara Lindgren 01/22/2000 PurposeThe protocol describes how to prepare selenomethionyl (Se-Met) protein using a regular E.coli strain. Selenium can be used for pha ...

Pectinase assayPectinases are actually a mixture of enzymes which along with others such as cellulase are widely used in the fruit juice industry where they are widely used to help extract clarify and ...

Purification of acidic phosphatase from mustard seedlingsPhosphate esters are widely distributed in any organism. Nucleic acids metabolic intermediates like glucose-6-phosphate energy-rich substrates ...

Radioiodination (by Jun Takagi6/16/2000)Purpose and backgroundsPrinciple of radioiodinationAddition of oxidizing reagents (such as chloramine-T or peroxidase+H202) converts I- to I+ or I3-. This highl ...

Column Purification of Demethylated SphingomyelinContributor: Suprya JayadevDate: Mar. 11 1991Packing column:1) To 20 g of 100-200 mesh Bio-Sil A silica gel add 80 mls of chloroform.2) Place a small p ...

Arachidonic Acid LabelingContributor: Suprya JayadevDate: November 4 19911) Grow cells to a density of 5-8 X 105 cells/ml in RPMI 1640 containing serum.2) Pellet cells and wash 1 time with room temper ...

Desalting Acidic GlycosphingolipidsContributor: Suprya JayadevDate: November 10 19931) Prepare a C-18 reverse phase prep-sep cartridge by washing with:a) 10 ml of methanolb) 20 ml of chloroform-methan ...

DGK AssayContributor: Suprya JayadevDate: Nov. 10 1995Buffers:- 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 ...

DGK Membrane PreparationContributor: Suprya JayadevDate: Dec. 10 1992Reagents:Bacterial strainE. coli N4830/pJW10LB amp media50 µg/ml ampicillinHigh salt bufferfor 1 L50 mM KH2PO4 6.8 g150 mM KCl 11.1 ...

In vitro Sphingomyelinase Assay Contributor: Suprya JayadevDate: May 25 1993Reagents: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 ...

Glycosphingolipid analysisContributor: Suprya JayadevDate: Nov. 10 19931) Incubate cells with 1 µCi/ml of 3H-galactose for 72 hours.--- If treatment is for an extended period of time: treat in serum f ...

Lipoprotein Isolation - First weekAs insects have an open circulatory system the hemolymph can be simply collected through an incision in the body wall. Most conveniently you should cut a leg and let ...

Lipoprotein Analysis Week 2: ElectrophoresisIntroductionSDS polyacrylamide gel electrophoresis (SDS PAGE) will be used to assess the purification process and to determine the apparent molecular weig ...