化学生物学实验技术

Labeled Sphingomyelin SynthesisContributor: Suprya JayadevDate: Mar. 11 1993Synthesis reaction:1) Mix purified DMSM with cyclohexylamine and 14CH3I at a ratio of 1/1.1/1.3 in 5 ml of methanol.2) Allow ...

Liposome PreparationOBJECTIVE:Method for incorporating proteins into liposomes for liposome swelling assay or as an alternative antigen presentation method.REAGENTS:Dioleoyl Phosphatidyl CholineBuffer ...

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 final volume of 0.6 ml. Flush with nitrogen.Stopper t ...

Western Blot(免疫印迹法)主要包括以下4个基本步骤：n 样品制备n 电泳分离n 蛋白的膜转移n 免疫杂交与显色��蛋白检测溶液和试剂n 1X 磷酸盐缓冲液(PBS)n Modified RIPA buffer Tris-HCl: 50 mM pH 7.4 ; NP-40: 1% ;Na-deoxycholate: 0.25% ;NaCl: 1 ...

Modified from Spudich & Watt 1971 JBC 246:4866.1. Mix 20 ml buffer G with each gm of muscle acetone powder. Extract with stirring on ice for 30 min. Use a big stir bar or overhead stirrer. Buffer G: 0 ...

DescriptionFor protein concentration gas pressure is applied directly to ultrafiltration cell. Solutes above the membrane's molecular weight (MW) cut-off are retained in cell while water and solutes b ...

Phosphate esters are widely distributed in any organism. Nucleic acids metabolic intermediates like glucose-6-phosphate energy-rich substrates (AMP creatine phosphate) are some obvious examples. While ...

Purpose and backgroundsPrinciple of radioiodinationAddition of oxidizing reagents (such as chloramine-T or peroxidase+H202) converts I- to I+ or I3-. This highly reactive molecule attacks o-position o ...

Typical modification sites: RGG box or RXR sequence motifs R-arginine G-glycineX-any aminoacid.Enzymes catalysing protein arginine methylation: PRMT1 from rat and the humanhomologue HRMT1L2 human PRMT ...

This specfic protocol is the latest incarnation of peptide mapping procedures that have been developed here in the TVL/MBVL of the Salk Institute over the last 20 years. Wade Gibson developed the firs ...

1. Immunoprecipitate the protein and run it on a preparative gel. CNBr cleavage must be done with protein transferred to a nitrocellulose filter. Neither Immobilon nor Nylon can substitute. IMPORTANT: ...

Polyacrylamide gel electrophoresis is a widely used technique to separate proteins from biological samples. Moreover the development of two-dimensional (2D) gel electrophoresis has provided a tool for ...

Materials: Overlay/Rehydration buffer:125 mM Tris-HCl pH 6.8 1mM EDTA 0.1% SDS 1mM 2-b-mercaptoethanol30% glycerolsome Bromophenol blueFor 1 ml you will need0.35 ml of 2X buffer0.30 ml H20 0.30 ml gly ...

Extraction:1) Following spin save supernatant for analysis. Be extremely careful not to disturb the pellet since it is somewhat dispersed.2) Immediately upon removing supernatant from each tube add 3 ...

1) Remove 2 500 µl aliquots of supernatant into scintillation vials add scintillation fluid and count.2) Aliquot 1.6 mls of the remaining supernatant into each of two pyrex glass tubes.3) Add 6 mls of ...

1) 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 temperature PBS.3) Resuspend final pellet in an appropriate volume of prewarmed ...

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.- dilution buffer1 ml 0.5 M imidazol pH 6.62.5 ml 20 ...

Packing column:1) To 20 g of 100-200 mesh Bio-Sil A silica gel add 80 mls of chloroform.2) Place a small portion of glass wool at the base of the column.3) Pour gel solution into column and use a stir ...

1) Prepare a C-18 reverse phase prep-sep cartridge by washing with:a) 10 ml of methanolb) 20 ml of chloroform-methanol (2:1)c) 10 ml of methanold) 10 ml of methanol-1.6 M sodium acetate (1:1)2) Apply ...

IntroductionMilk fat measurement is a common task in the dairy industry because milk fat content is one factor that determines milk price and is necessary to know for casein/fat ratio normalization. I ...