WesternBlottingProtocol
实验概要The western blot (sometimes called the protein immunoblot) is a widely used analytical technique used to detect specific proteins in the given sample of tissue homogenate or extract. It uses gel electrophoresis to separate native proteins by 3-D structure or denatured proteins by the length of the polypeptide. The proteins are then transferred to a membrane (typically nitrocell......阅读全文
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
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
Radioiodination－of－protein
Radioiodination (by Jun Takagi,6/16/2000) Purpose and backgrounds Principle of radioiodination Addition of oxidizing reagents (such as chloram
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
Protein－Electrophoresis
DefinitionAmino acids, nucleotides, polypeptides, and other compounds in a colloidal state can be separated by the application of external voltages wh
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 i
Protein－Kinase－A－at－the－Centrosome
Protein kinase A regulatory subunit RIIalpha (PKA-RIIa) is tightly bound to centrosomal structures during interphase through
Acetone－precipitation－of－protein
This procedure is suitable for recovering proteins from most aqueous solvents and from SDS containing buffers. It is not recommended for proteins dis
BIURET－PROTEIN－ASSAY
BIURET PROTEIN ASSAY MATERIALS Biuret Reagent Bovine serum albumin (BSA) Spectrophotometer and tubes PROCEDURE Prepare standard d
Preparing－a－Selenomethionyl－Protein
Purpose The protocol describes how to prepare selenomethionyl (Se-Met) protein using a regular E.coli strain. Selenium can be used for phase determi
Protein－arginine－methylation
Typical modification sites: RGG box or RXR sequence motifs R-arginine, G-glycine, X-any aminoacid. Enzymes catalysing protein arginine methylation: P
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
Protein－A－Purification－of－Antibody
1. Reagents(1) Affi-gel Protein-A Agarose (BioRad #153-6153)(2) MAPS II Binding Buffer (BioRad # 153-6161)(3) 0.314 g/ml diH2O(4) MAPS II Elution Buff
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－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－Determination
Bradford – Protein DeterminationIntroductionA rapid and accurate method for the estimation of protein concentration. The technique is simpler, faster
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
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,
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-
Lowry－–－Protein－Determination
Lowry – Protein Determination(From Protein Protocols on CD-ROM Humana Press, 1998 - Section 1-2 The Lowry Method for Protein Quantitation Jakob H. Wat
Basic－Protein－Chemistry－Techniques
Coomassie Blue Stain: (for gels) 1) Combine 225 ml Methanol with 225 ml ddH2O. 2) Add 0.5 grams of Coomassie Blue. 3) Just before use, add 50 ml
Mechanism－of－Protein－Import－into－the－Nucleus
Nuclear pore complexes (NPCs) are large proteinaceous assemblies that provide the only known portals for exchang
Basic－Protein－Chemistry－Techniques
实验概要 Basic Protein Chemistry Techniques 实验步骤 Coomassie Blue Stain: (for gels) 1) Combine 225 ml Methanol with 225 ml ddH2O. 2) Add 0.5 grams of
Angiotensin－Protein－Kinase－Assay
James Hardwick's angiotensin assay protocol This specific procedure was developed to assay the activity of the Lck kinase expressed from a retroviral
Biorad－Protein－Assay:－Bradford
Biorad Protein Assay: BradfordStandards: 1 mg/ml BSA stock- dilute 1:10 to get 0.1 mg/ml BSAAdd To get H-2O20 µl 2 µg/ml 780 µl40 µl 4 µg/ml 760 µl60
Bradford－Protein－Concentration－Assay
Bradford Protein Concentration Assayversion 01/07/2001Abbreviations:mcg = microgramsmcL = microlitersBSA = bovine serum albuminO.D. = optical densityd
Protein－G－Purification－of－Antibodies
1. Reagent and Materials(1) Hi-Trap Protein G Column (Pharmacia Biotech #17-0404-01)(2) 20 mM Sodium Phosphate Buffer, pH 7.01.084 g NaH2PO4, anhydrou
Protocol－for－Protein－Extraction－for－proteomics
Protocol for Protein Extraction10 % w/v TCA/ acetone/ 0.07 % v/v -MercaptoethanolPlant cells are rich in compounds that interfere with the 2DE separat
Protein－Immunolocalization－in－Maize－Tissues
The analysis of gene expression at transcript and at protein level is of outstanding importance in plant developmental biology. Proteins can be loc
Protein－Expression－and－Purification－Protocol
Step 1: Transform appropriate DNA plasmid into BL21(DE3) E. coli cells. These cells must be competent. (Protocol for how to make competent cells.)a) T