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......閱讀全文
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、實驗試劑 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
實驗概要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
信號傳導
Cytokine Bioassays?(eBioscience)Biological activity of cytokines and their concentrations are commonly measured by cellular proliferation of primary c
信號傳導
Cytokine Bioassays?(eBioscience)Biological activity of cytokines and their concentrations are commonly measured by cellular proliferation of primary c
四唑鹽（MTT）比色實驗——MTT法
實驗方法原理活細胞的線粒體中的琥珀脫氫酶能使外源性的MTT還原為難溶性的藍紫色結晶物并沉積在細胞中，而死細胞無此功能。二甲基亞砜能溶解細胞中的紫色結晶物，用酶聯免疫檢測儀在490? nm 波長處測定其光吸收值，可間接反映細胞數量。在一定細胞數范圍內，MTT結晶物形成的量與細胞數成正比。?實驗材料細胞
小鼠脾臟T細胞和人PBMCs的體外T細胞活化實驗（二）
材料1X 無菌 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實驗心得
MTT實驗是檢測細胞活力的實驗方法，由于細胞活力與細胞數呈正相關，因此也常常用來檢測細胞的增殖情況。MTT的原理：活細胞有琥珀酸脫氫酶，將MTT還原成棕褐色沉淀。由于一般介紹園子里已經很多，筆者將自己的心得按照實驗流程與大家交流交流。1、培養好細胞點板。養細胞沒啥好說的，如果不知道細胞如何養，那就看