ProteinImmunolocalizationinMaizeTissues
The analysis of gene expression at transcript and at protein level is of outstanding importance in plant developmental biology. Proteins can be localized with subcellular resolution by immunolocalization using specific antibodies or generating reporter lines carrying the specific protein fused to a fluorescent protein. Immunolocalization is particularly suitable to confirm the expr......阅读全文
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
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
AgrobacteriumMediated－Maize－Transformation:－Immature－Embryos－Versus－Callus
Transformation with Agrobacterium tumefaciens is the preferred method for delivery of transgenes into a wide range of plant species including maiz
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
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 in
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
BIURET－PROTEIN－ASSAY
BIURET PROTEIN ASSAYMATERIALSBiuret ReagentBovine serum albumin (BSA)Spectrophotometer and tubesPROCEDUREPrepare standard dilutions of BSA containing
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
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
Protein－purification;－actin
Protein purification; actin Overview ACTINThe most abundant muscle and non-muscle cytoskeletal protein. MW 42 kDa, 374/375 amino acids; various
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
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 diss
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
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
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－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,
Bradford－–－Protein－Determination
Bradford – Protein DeterminationIntroductionA rapid and accurate method for the estimation of protein concentration. The technique is simpler, faster
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
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-
In－Vitro－Protein－Ubiquitination－Assay
Ubiquitination is one of the most important posttranslational modifications in all eukaryote organisms. Ubiquitin-activating enzyme (E1), ubiquiti
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
Mechanism－of－Protein－Import－into－the－Nucleus
Nuclear pore complexes (NPCs) are large proteinaceous assemblies that provide the only known portals for exchanging macromolecules between the nucleus
Angiotensin－Protein－Kinase－Assay
James Hardwick's angiotensin assay protocolThis specific procedure was developed to assay the activity of the Lck kinase expressed from a retrovir
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 acet
Coupling－Antibodies－to－Protein－A－or－G
1. use 2 mg of antibody per ml wet beads (use appropriate antibody/protein A or G combination).2. mix antibodies with beads and bind at room temperatu
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
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