InVitroProteinUbiquitinationAssay
Ubiquitination is one of the most important posttranslational modifications in all eukaryote organisms. Ubiquitin-activating enzyme (E1), ubiquitin-conjugating enzyme (E2), and ubiquitin ligase (E3) are the three key enzymes in this process. To detect the specificity between E2 and E3 or enzyme–substrate relationship between E3 and a substrate protein, ubiquitination activity needs to be determined. This pr......阅读全文
In－Vitro－Protein－Ubiquitination－Assay
Ubiquitination is one of the most important posttranslational modifications in all eukaryote organisms. Ubiquitin-activating enzyme (E1), ubiquiti
In－Vivo－Ubiquitination－Assay－by－Agroinfiltration
The ubiquitination/proteasome system is involved in nearly all plant signaling processes. Many signaling components are degraded by the 26S protea
In－Vitro－Fertilization
When we first started using X. tropicalis, in vitro fertilization had an extremely poor efficiency. However, with the careful selection of a mature ma
In－vitro－growth－of－seedlings
sterilisation of seeds: rinse with 70% EtOH for 30 sec put in 1% bleach (sodium hypochlorite, supplemented by a few drops of Tween-20) for 5 mi
In－vitro－Sphingomyelinase－Assay
Reagents:Lysis buffer25 mM Tris-HCl, pH 7.45 mM EDTA1 mM ATP20 µg/ml CLAP1 mM PMSFBuffer A10 mM MgCl20.2 M Tris-HCl, pH 7.40.2 % Triton X-100Buffer B0
InVitro－Adipocytes－Differentiation
IntroductionObesity is a significant clinical problem that contributes to life-threatening diseases such as diabetes and atherosclerosis. With an incr
In－vitro－culture－of－embryonic－lungs
In vitro culture of embryonic lungsfrom Hogan LabIsolation of Lung Bud EndodermWhat you need:E11-12 mouse embryosDMEM with 5% fetal bovine serumpetri
体外重组（in－vitro－recombination）
载体与外源DNA分子体外重组时，如何选择优化连接条件以达到最高的重组率。因此有必要根据影响连接效率的因素综合考虑连接条件。影响连接效率的因素很多，如反应温度、插入片段和载体之间的摩尔比、DNA末端性质、反应时间、ATP浓度等。1. 反应温度是比较重要的影响因素。因为连接酶的最适反应温度为37℃，
In－Vitro－T－Cell－Activation
In Vitro T Cell ActivationIntroductionMature T cells recognize and respond to the antigen/MHC complex through their antigen-specific receptors (TCR).
Coating－of－Platelets－with－Antibody－in－vitro
OUTLINEAntibody-coated platelets (opsnized) may be used in the subsequent thrombophagocytosis assay. PROTOCOLResuspend 1.6x10^8 of CMFDA-labeled plate
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
Invitro－Phagocytosis－Assay－of－Macrophages
IntroductionThe term phagocytosis itself describes its mean phage = engulfment; cytosis: cell process. In other words, phagocytosis is the cellular pr
In－Vitro－Conservation－and－Cryopreservation－of－Ornamental－Plants
Today, the conservation of ornamental germplasm can take advantage of innovative techniques which allow preservation in vitro (slow growth storage
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
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
Preparation－of－Luciferin－for－In－Vitro－and－In－Vivo－Bioluminescent－Assays
Preparation of Luciferin forIn Vitro Bioluminescent AssaysMaterials• D-Luciferin Firefly, potassium salt, 1.0 g /vial(Caliper Life Sciences Part Numbe
Preparation－of－Luciferin－for－In－Vitro－and－In－Vivo－Bioluminescent－Assays
实验概要Reagent for immunoassay, ligand binding assay and ligand receptor assay in which luciferin is covalently bonded to a molecule having biological
In－vitro－Assessment－of－Metabolic－－in－Suspension－Cryopreserved－Hepatocytes
实验概要BackgroundThe pharmaceutical and biotechnology industry’s goal is to discover therapeutic agents that are both safe and effective at treating or
Preparation－of－Luciferin－for－In－Vitro－and－In－Vivo－Bioluminescent－Assays
实验概要Reagent for immunoassay, ligand binding assay and ligand receptor assay in which luciferin is covalently bonded to a molecule having biological
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
Betagal－staining－of－eukaryotic－cells－in－vitro
(Modification of methods of Dr. Seong-Seng Tan and Promega's "Protocols and Applications Guide")Cells previously transfected with a lac Z construc
A－novel－in－vitro－3dimensional－angiogenesis－model
1. Human microvascular endothelial cells (HMVECs) with primary cell kits were cultured on collagen type I-coated dishes to 80% confluency, then ov
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－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
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
BIURET－PROTEIN－ASSAY
BIURET PROTEIN ASSAYMATERIALSBiuret ReagentBovine serum albumin (BSA)Spectrophotometer and tubesPROCEDUREPrepare standard dilutions of BSA containing
Bradford－–－Protein－Determination
Bradford – Protein DeterminationIntroductionA rapid and accurate method for the estimation of protein concentration. The technique is simpler, faster