CriticalAppraisaloftheMTTAssayinthePresenceofRottlerin3
LDH assay LDH assay was performed in culture medium of untreated confluent cells by using a commercial kit (Sclavo Diagnostics, Siena) based on the transformation of pyruvate to lactate by LDH, at pH 7.5, in the presence of NADH coenzyme. The transformation of NADH to NAD+ is accompanied by a decrease in absorbance (A) at 340 nm, which correlates with the LDH activity. The change of absorbance, in th......阅读全文
BIURET－PROTEIN－ASSAY
BIURET PROTEIN ASSAY MATERIALS Biuret Reagent Bovine serum albumin (BSA) Spectrophotometer and tubes PROCEDURE Prepare standard d
In－vitro－Sphingomyelinase－Assay
Reagents: Lysis buffer 25 mM Tris-HCl, pH 7.4 5 mM EDTA 1 mM ATP 20 µg/ml CLAP 1 mM PMSF Buffer A 10 mM MgCl2 0.2 M Tris-HCl, pH 7.4 0.2 % Triton X
ELISA－Inhibition－Assay
ELISA Inhibition AssaySensitize a 96-well microtiter plate with purified antigen.Prepare a solution of the purified antigen of interest in phosphate b
cell－proliferation－assay
cell proliferation assaybefore start:thaw cells from liquid nitrogen, grow in 75cc flask (T75) in Fischer's medium MM (maintenance medium) until c
HISTONE－KINASE－ASSAY
PROTOCOLTo 1.5 mL eppendorf tubes add:200 µg of protein extract (see Western blot protocol for protein sample preps)q.s. to 300 µL with RIPA (with pro
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
Glycolipid－Binding－Assay
Glycolipid Binding AssaySource: Contributed by Pingsunjim, Paller’s LabAbstract: This protocol can be used for the detection of glycolipids binding to
DNA－methyltransferase－Assay
Methylated CpG island Amplification Protocol written by Minoru Toyota2. Materials2.1. MCARestriction enzymes SmaI, XmaIT4 DNA ligaseTaq DNA polymerase
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,
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
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-
Crystal－Violet－Assay
This is a simple assay useful for obtaining quantitative information about the relative density of cells adhering to multi-well cluster dishes. The dy
Migration－Assay－Protocol
Materials to be prepared beforehand:1) FBS free medium2) 10% FBS medium3) Cell migration filter insert ( Transwell®, 12mm Diameter, 12 μm Pore Size.)P
Adhesion－Assay－Protocol
Materials to be prepared beforehand:1) Washing Buffer--0.1% BSA in medium (DMEM or RPMI)2) Blocking Buffer--0.5% BSA in medium (DMEM or RPMI)3) Lamini
Wound－healing－assay
The wound healing assay allows the researcher to study cell migration and cell interactions. In some cases also single cell migration can be analyzed.
Tube－formation－assay
DescriptionThis is a fast and easy assay to test the angiogenic/anti-angiogenic properties of molecules. As compared to other angiogenesis assays, suc
Cell－Viability－Assay
Dye exclusiona cell suspension is mixed with trypan blue and examined by low-power microscopyMaterialscellsPBSM3hemocytometer0.4 % trypan blue in PBSm
Soft－Agar－Assay
Soft Agar AssayMake 0.6% media-agar mix for the bottom layer. To make 0.6% agar mix the following components (this makes 200 ml):2X DME 100 mlI
Glucosamine－Rapid－Assay
Glucosamine Rapid AssayMRTHOD:Place sample (containing 0.5 - 10 µg GlcN) in a Pyrex screw capped tube.Add HCl to a final concentration of 2N and a fin
Leaf－GUS－Assay
一、实验试剂 GUS Buffer (500 ml) 2.0478 g Na2HPO4 1.2688 g NaH2PO4 (=50 mM NaPi pH7.0) 10 ml 0.5 M EDTA (=10 mM) 0.5 g Triton X-100 0.5 g N-L
Leaf－GUS－Assay
实验概要a protocol for Leaf GUS Assay This protocol is for small samples (usually single leaf from 21DAI plants), scale up for larger samplesAs there are
Pheromone－Halo－Assay
-Use sterile technique and sterile solutions throughout this method.-1. Grow a starter culture at 30 C with shaking (250 rpm) until it reaches saturat
信号传导
Cytokine Bioassays (eBioscience)Biological activity of cytokines and their concentrations are commonly measured by cellular proliferation of primary c
信号传导
Cytokine Bioassays (eBioscience)Biological activity of cytokines and their concentrations are commonly measured by cellular proliferation of primary c
四唑盐（MTT）比色实验——MTT法
实验方法原理活细胞的线粒体中的琥珀脱氢酶能使外源性的MTT还原为难溶性的蓝紫色结晶物并沉积在细胞中，而死细胞无此功能。二甲基亚砜能溶解细胞中的紫色结晶物，用酶联免疫检测仪在490 nm 波长处测定其光吸收值，可间接反映细胞数量。在一定细胞数范围内，MTT结晶物形成的量与细胞数成正比。 实验材料细胞
小鼠脾脏T细胞和人PBMCs的体外T细胞活化实验（二）
材料1X 无菌 PBSAnti-human CD3:Clone OKT3 (Functional Grade， eBioscience Cat. No. 16-0037) or Clone HIT3a (Functional Grade， eBioscience Cat. No. 16-0039)R
The－Impact－of－Harmo...－（二）
In addition, proficiency panel projects have demonstrated that even after an experiment was done and spots were counted, considerable variation can
COMPARISION－OF－DIFFERENT－PROTEIN－DETERMINATION－METHODS
COMPARISION OF DIFFERENT PROTEIN DETERMINATION METHODSCompanyMethodDetection RangeApplications -CompatibilityAssay protocolPrecautions-InterferencesA
T－cell－Activation－Protocol
IntroductionMature T cells recognize and respond to the antigen/MHC complex through their antigen-specific receptors (TCR). The most immediate consequ
MTT实验心得
MTT实验是检测细胞活力的实验方法，由于细胞活力与细胞数呈正相关，因此也常常用来检测细胞的增殖情况。MTT的原理：活细胞有琥珀酸脱氢酶，将MTT还原成棕褐色沉淀。由于一般介绍园子里已经很多，笔者将自己的心得按照实验流程与大家交流交流。1、培养好细胞点板。养细胞没啥好说的，如果不知道细胞如何养，那就看