Roche公司的RNaseProtectionAssay(RPA)protocol

Roche公司的RNase Protection Assay (RPA) Using DIG-Labeled RNA Probes下载网址:http://www.roche-applied-science.com/PROD_INF/BIOCHEMI/no1_03/PDF/p22_23.pdf还有一份protocolhttp://www.ualberta.ca/PERINATAL/facilities/pdf/RNase%20Protection%20Assay.pdf可能出现的问题及应对措施1.No Discrete Bands, Only Smears--RNA degraded, or the the RNase Digestion was too harsh.Try reducing the concentration of RNase A to 1 mg/ml or digest for a shorter period......阅读全文

ELISPOT-Protocol

实验概要The  Elispot (Enzyme Linked Immuno-Spot) assay provides an effective method  of measuring the antibody or cytokine production of immune cells on t

ELISPOT-Protocol

实验概要The  Elispot (Enzyme Linked Immuno-Spot) assay provides an effective method  of measuring the antibody or cytokine production of immune cells on t

ELISA-protocol

ELISA protocol:1.取5-10ul BMMY表达上清用0.05M NaHCO3稀释到100ul铺ELISA板,37度或室温振荡大于1小时。注意一定要做一个GS115空菌株表达上清作为阴性对照,最好还找一个带有histag的蛋白作为阳性对照。2.TPBS洗板3次,方法:倒掉铺板液,倒置于

Immunoblot-Protocol

This method was successful in our lab using prostate tissue and for our specific objectives. Investigators must be aware that they will need to tailor

NAi-protocol

siRNA protocolsOur current strategy with siRNA is to synthesis relatively small amounts enzymatically and use these to test for efficiency by western

ELISPOT-protocol

实验概要The procedure  below is a general guideline procedure for ELISPOT. Abcam ELISPOT kits  have been designed for detection of various cytokines and g

PCR-protocol

PCR reactionProtocol for 50µl reaction - adjust amounts if necessary, for a 20µl reaction use the same volumes of primer and dNTP-mix, but adjust the

RLGS-protocol

A. Preparation of DNA SolutionIn the case of rice, for example    This method may be appllicable for many grass species and some other plants.        

RNAi-protocol

 siRNA protocolsOur current strategy with siRNA is to synthesis relatively small amounts enzymatically and use these to test for efficiency by western

Actin-Capture-Assay

David AmbergDialyze purified GST fusion proteins and actin into PBS + 1mM MgCl2 .Mix 5ug actin into 50ul total volume binding buffer.Mix 5ug GST-fusio

Assay-for-the-Micrococcal-Nuclease

Method: Essentially that described by Heins et al. (1966) based upon the release of acid soluble oligonucleotides following nuclease digestion of DNA.

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

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

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