MTTAssay
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 PBS and filter sterilise.5 hours before the end of the incubation add 20ml of MTT solution from step one to each well containing cells.Incubate the plate at 37篊 for 5 hours.Remove media with needle and syringe.Add 200m l of DMSO to each well and pipette up an......阅读全文
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-
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
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
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
BIURET－PROTEIN－ASSAY
BIURET PROTEIN ASSAYMATERIALSBiuret ReagentBovine serum albumin (BSA)Spectrophotometer and tubesPROCEDUREPrepare standard dilutions of BSA containing
DNA－methyltransferase－Assay
Methylated CpG island Amplification Protocol written by Minoru Toyota2. Materials2.1. MCARestriction enzymes SmaI, XmaIT4 DNA ligaseTaq DNA polymerase
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
Assay－of－Phospholipase－A－Activity
Phospholipases of the A type constitute a large family of esterases that catalyze the hydrolysis of the fatty acid ester bonds in phospholipids an
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
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
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,
Cell－Viability－Assay
Dye exclusiona cell suspension is mixed with trypan blue and examined by low-power microscopyMaterialscellsPBSM3hemocytometer0.4 % trypan blue in PBSm
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
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
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.
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
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
Glycolipid－Binding－Assay
Glycolipid Binding AssaySource: Contributed by Pingsunjim, Paller’s LabAbstract: This protocol can be used for the detection of glycolipids binding to
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
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
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
Needle－Assay－for－Chemotaxis
Devreotes Lab, John Hopkins Medical Institutions http://www.hopkinsmedicine.org/cellbio/devreotes/needle.htmEquipment and chemicalsZeiss inverted micr
MTT法实验原理与MTT－溶液的配制方法
通常，此法中的mtt浓度为5mg/ml。因此，可以称取mtt0.5克，溶于100ml的磷酸缓冲液(pbs)或无酚红的培养基中，用0.22μm滤膜过滤以除去溶液里的细菌，放4℃避光保存即可。在配制和保存的过程中，容器最好用铝箔纸包住。需要注意的是，mtt法只能用来检测细胞相对数和相对活力，但不能测定细
MTT法实验原理与MTT溶液的配制方法
通常，此法中的mtt浓度为5mg/ml。因此，可以称取mtt0.5克，溶于100ml的磷酸缓冲液(pbs)或无酚红的培养基中，用0.22μm滤膜过滤以除去溶液里的细菌，放4℃避光保存即可。在配制和保存的过程中，容器最好用铝箔纸包住。需要注意的是，mtt法只能用来检测细胞相对数和相对活力，但不能测定细
mtt实验中细胞处于什么状态时加入mtt
要进行预实验检测其贴壁率、MTT，尽量无菌操作，细胞会由增殖期渐渐趋向G0期而趋于静止，才能保证MTT结晶形成数量与细胞数呈的线性关系。要根据自己的实际情况调整。用含15%胎牛血清培养液培养细胞时，太少观察不到差异。一定要多看文献，不能测定细胞绝对数，以保证培养终止致细胞过满，可能你用的时间和浓度根
MTT法实验原理与MTT溶液的配制方法
通常，此法中的mtt浓度为5mg/ml。因此，可以称取mtt0.5克，溶于100ml的磷酸缓冲液(pbs)或无酚红的培养基中，用0.22μm滤膜过滤以除去溶液里的细菌，放4℃避光保存即可。在配制和保存的过程中，容器最好用铝箔纸包住。需要注意的是，mtt法只能用来检测细胞相对数和相对活力，但不能测定细
MTT法实验原理与MTT－溶液的配制方法
原理：活细胞中脱氢酶能将四唑盐还原成不溶于水的蓝紫色产物甲臜，并沉淀在细胞中，而死细胞没有这种功能。DMSO能溶解沉积在细胞中蓝紫色结晶物，溶液颜色深浅与所含的formazan量成正比。再用酶标仪测定OD值。
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
Cytokine－Bioassays
Cytokine BioassaysIntroductionBiological activity of cytokines and their concentrations are commonly measured by cellular proliferation of primary cel