生物化学技术

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 ...

Lipoprotein Analysis -week 3During this week you will analyze the SDS PAGE and determine the protein content in the lyophilized fraction of lipoproteins gravimetrically. You will also extract the lipi ...

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 ...