CyanogenBromidedigestionofprotein
1. Proteins are TCA precipitated and washed with acetone, then dried.2. The CNBr should be brought to room temperature in the hood and used ONLY in the hood. Extremely toxic!3. Dissolve a small amount of CNBr in 70% TFA, the exact amount is not critical. One or two crystals should be fine, although weighing this reagent is NOT recommended. Place the utensil used in dilute bleach to neutralize the residual CNBr.4. Red......阅读全文
Cyanogen－Bromide－digestion－of－protein
1. Proteins are TCA precipitated and washed with acetone, then dried.2. The CNBr should be brought to room temperature in the hood and used ONLY in th
Ethidium－bromide
Introduction Ethidium bromide (EtBr), 3,8-diamino-5-ethyl-6-phenylphenanthridinium bromide, is commonly used as a non-radioactive marker for identify
Ethidium－Bromide－Decontamination
N.B.: Ethidium bromide is a powerful mutagen. Protective gloves should be worn at all times when handling solutions containing ethidium bromide.Decont
Restriction－digestion
Restriction digestionRestriction enzyme digestions are performed by incubating double-stranded DNA molecules with an appropriate amount of restriction
Partial－Endonuclease－Digestion
Partial Endonuclease DigestionPrepare a 100 µl reaction mixture containing the DNA of interest in the appropriate 1X restriction enzyme buffer. Divide
Two－dimensional－peptide－mapping
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
Decontamination－of－Ethidium－Bromide－Solutions－and－Surfaces
WARNING: EtBr is toxic and mutagenic. Hypophosphorus and its solutions are\corrosive. Decontamination solution gives off a small amount of nitrogendio
Restriction－Enzyme－Digestion－of－DNA
Materials:10X restriction enzyme buffer (see manufacturer's recommendation)DNAsterile waterrestriction enzymephenol:chloroform (1:1)Add the follow
Antibody－Purification－using－Protein－A,－Protein－G,－or－Protein－L－Agarose
实验概要This protocol is designed as a quick purification method for antibodies from mammalian sera, ascites, and cell culture supernatants主要试剂 Protein
Antibody－Purification－using－Protein－A,－Protein－G,－or－Protein－L－Agarose
实验概要This protocol is designed as a quick purification method for antibodies from mammalian sera, ascites, and cell culture supernatants. It should
James－Hardwick－CNBr－Cleavage－Procedure
1. Immunoprecipitate the protein and run it on a preparative gel. CNBr cleavage must be done with protein transferred to a nitrocellulose filter. Neit
10mg/ml的溴化乙锭（ethidium－bromide）
小心称取1g溴化乙锭，转移到广口瓶中，加100ml水，用磁力搅拌器搅拌直到完全溶解。用铝箔包裹装液管，于4℃贮存。
Ingel－digestion－of－proteins－for－peptide－fingerprint－mapping
Polyacrylamide gel electrophoresis is a widely used technique to separate proteins from biological samples. Moreover, the development of two-dimension
Protein－Electrophoresis
DefinitionAmino acids, nucleotides, polypeptides, and other compounds in a colloidal state can be separated by the application of external voltages wh
Protein－Crystallization
Background:Proteins, like many molecules, can be prompted to form crystals when placed in the appropriate conditions. In order to crystallize a protei
Radioiodination－of－protein
Radioiodination (by Jun Takagi,6/16/2000)Purpose and backgroundsPrinciple of radioiodinationAddition of oxidizing reagents (such as chloramine-T or pe
The－Determination－of－Proteinprotein－Interactions－by－the－Matingbased－...
Dynamic and reversible protein–protein interactions have a pivotal function in all living cells. For instance, protein–protein interactions are in
InGel－Digestion－of－Proteins－Separated－byPolyacrylamide－Gel－Electrophoresis
1. Excision of protein bands (spots) from polyacrylamide gelsRinse the gloves you use with water to avoid traces of dust in your sample.Rinse the gel
亲和层析实验技术方法
INTRODUCTIONThis protocol describes a method for removing antibodies that react with bacterially encoded proteins by passing a crude preparation of im
Protein－Assay－(Spectrophotometer)
Protein Assay (Spectrophotometer)Use BSA (bovine serum albumin) 1mg/ml stock solution (1ml Eppendorf tubes) for standard curve.Place 0, 2, 5, 10, 15,
Eukaryotic－protein－translation
The scanning translation initiation model suggests that 40S ribosomal subunit preloaded with factors bind to the 5’ end of the mRNA near the cap. The
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-
Protein－purification;－actin
Protein purification; actin Overview ACTINThe most abundant muscle and non-muscle cytoskeletal protein. MW 42 kDa, 374/375 amino acids; various
Protein－arginine－methylation
Typical modification sites: RGG box or RXR sequence motifs R-arginine, G-glycine,X-any aminoacid.Enzymes catalysing protein arginine methylation: PRMT
Protein－Kinase－A－at－the－Centrosome
Protein kinase A regulatory subunit RIIalpha (PKA-RIIa) is tightly bound to centrosomal structures during interphase through interaction with the A-ki
Protein－Staining－Procedures
This method was successful in our lab using prostate tissue and for our specific objectives. Investigators must be aware that they will need to tailor
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
Bradford－–－Protein－Determination
Bradford – Protein DeterminationIntroductionA rapid and accurate method for the estimation of protein concentration. The technique is simpler, faster
BIURET－PROTEIN－ASSAY
BIURET PROTEIN ASSAYMATERIALSBiuret ReagentBovine serum albumin (BSA)Spectrophotometer and tubesPROCEDUREPrepare standard dilutions of BSA containing
Preparing－a－Selenomethionyl－Protein
PurposeThe protocol describes how to prepare selenomethionyl (Se-Met) protein using a regular E.coli strain. Selenium can be used for phase determinat