生物化学技术

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 ...

Polygalacturonase assayThis 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 of these tomatoes have been ...

Polyphenoloxidase (catechol oxidases) assayBrowning of the cut surface of some fruits and vegetables is due the presence of a group of enzymes called polyphenoloxidases. These enzymes are released by ...

Protease assayIn certain fruits such as pineapples and mangoes the flesh contains protein-digesting enzymes (proteases). These may play a part in helping to soften the fruit tissues as the fruit ripen ...

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 ...

AA & Metabolite Quantitation of Cell Pellets Post-AA Labeling Contributor: Suprya JayadevDate: November 4 1991Extraction:1) Following spin save supernatant for analysis. Be extremely careful not to di ...

AA & Metabolite Quantitation of Media Post-AA Labeling Contributor: Suprya JayadevDate: November 4 19941) Remove 2 500 µl aliquots of supernatant into scintillation vials add scintillation fluid and c ...

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 ...

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

Lipid analysisEXTRACTION OF LIPIDS FROM LIPOPHORIN Since lipids are hydrophobic they are better soluble in organic solvents than in water. Because the lipids are buried into protein particles simple ...

Cellulase assays Cellulase enzymes show activity during the ripening of some fruits where their effects on cell walls results in softening of the fruit. In cases of programmed cell death such as the f ...

Enzyme Kinetics assay of the WT To assay 17 b-HSD activity in lysates cells were harvested 48h after transfection using PBS Enzyme Free Cell Dissociation Solution ( specialty Media Inc. Lavellette NJ) ...

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 ...

Methods for the Detection of D-Amino-Acid Oxidase Ryuichi Konnoundefined1 Department of Microbiology. Dokkyo University School of Medicine Mibu Tochigi 321-0293. Japan. * To whom correspondence should be add ...

重组蛋白质的表达、纯化、复性和定量按Qiagen公司的操作手册进行，具体步骤如下。一、重组蛋白质的诱导表达1.挑取转化有质粒的单菌落，接种于3ml 选择性LB液体培养基中，37 oC，250 rpm/min振摇培养过夜。2.次日将培养过夜的菌液500 μl再接种于10 ml（1:20）选择性LB液体培养基中，37 oC，250 rpm/min振摇培养至光密度（OD600=0.6）时，取1 ml样本 ...

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 ...

PVDF膜上蛋白的可逆染色 Western杂交时，为确认蛋白是否转至PVDF膜上，可用下列方法对膜上蛋白进行可逆性染色。1. 氨基黑染色染液：0.5% Amido Black (w/v) 25% isopropanol (v/v) and 10% acetic acid. 染色：将PVDF膜置于染液中染色数钟，ddH2O 脱色。2. 考马氏亮蓝染色染液：0.1% Coomassie ...

方法：1、粗酶液的制备用电子天平分别称取粗脂肪酶0.010 g、0.020 g 和0.030 g， 用蒸馏水溶解并定容至100 mL， 配成浓度分别为0.01%、0.02% 和0.03%的粗酶液。2、实验设计本实验以10 mL色拉油为底物，以酶用量、水解温度、反应时间为因素，通过酸价的测定选定其水解的最佳条件。3、酸价的测定酸价是指中和1 mol游离脂肪酸所需NaOH的毫克数， 它用于衡量油脂的水 ...

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 fewer or more cells depending on your needs. ...