InvitroSphingomyelinaseAssay
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 B0.2 M sodium acetate, pH 5.00.2 % Triton X-100Buffer C0.2 M Tris-HCl, pH 7.40.2 % Triton X-100Isolating cellular enzyme1) Grow 5 x 108 cells under regular growth conditions.2) Pellet cells and wash one time with ice cold PBS.3) Resuspend pellet in 10 ml of ice cold lysis......阅读全文
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
In－Vitro－Protein－Ubiquitination－Assay
Ubiquitination is one of the most important posttranslational modifications in all eukaryote organisms. Ubiquitin-activating enzyme (E1), ubiquiti
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－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－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
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
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℃，
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－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
Motility－Assay
DescriptionVarious phenotypic characteristics are requiredfor a cancer cell to successfully complete the metastaticcascade. Among these, acquisition o
DGK－Assay
Buffers:- 2X buffer10 ml 0.5 M imidazol, pH 6.60.21 g LiCl1.25 ml 1 M MgCl21.0 ml 0.1 M EGTA, pH 6.6--> Bring volume up to 50 ml with distilled water.
Protease－assay
实验概要 In certain fruits, such as pineapples and mangoes, the flesh contains protein-digesting enzymes (proteases). These may play a part in
Chemotaxis－Assay
PurposeThe purpose of a chemotaxis assay is to determine whether your protein or small molecule of interest has chemotactic activity on a specific cel
Pectinase－assay
Pectinases are actually a mixture of enzymes, which, along with others such as cellulase, are widely used in the fruit juice industry where they are w
Protease－assay
In certain fruits, such as pineapples and mangoes, the flesh contains protein-digesting enzymes (proteases). These may play a part in helping to softe
Bradford－Assay
Bradford AssayThe bradford dye-binding assay is a colorimetric assay for measuring total protein concentration. It involves the binding of Coomassie B
TUNEL－assay
PROTOCOL:•Deparaffinize and rehydrate slides:3 x 3´ Xylene3 x 2´ 100% ethanol1 x 2´ 95%, 80%, 70% ethanol (each)1 x 5´ 1x PBS•Microwave antigen retrie
Phosphate－Assay
1. Make standards using sodium phosphate at the following uM concentrations: 0, 2, 5, 7, 10, 20, 40, 60, and 80. Use the screw top glass tubes.2. Dry
Bradford－Assay
The bradford dye-binding assay is a colorimetric assay for measuring total protein concentration. It involves the binding of Coomassie Brilliant blue
MTT－Assay
This procedure is for cells in 96 well plates, if larger plates are used then adjust volumes accordingly.1 Make a solution of 5mg/ml MTT dissolved in
Aspartate－Assay
实验概要The Aspartate Assay Kit provides a simple, convenient assay to measure aspartate in a variety of samples. In the assay, aspartate is converted
Polygalacturonase－assay
This 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 o
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
DNA－methyltransferase－Assay
Methylated CpG island Amplification Protocol written by Minoru Toyota2. Materials2.1. MCARestriction enzymes SmaI, XmaIT4 DNA ligaseTaq DNA polymerase
Noble－Agar－Assay
DescriptionCancer cells do not show anchorage and contact inhibition of growth. To assess the anchorage and contact independent growth of cells, noble