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

Roche公司的RNase-Protection-Assay-(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

The-ribonuclease-protection-assay-(RPA)

The ribonuclease protection assay (RPA) is a highly sensitive and specific method for the detection of mRNA species. The assay was made possible by th

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

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

Xenograft-Tumor-Assay-Protocol

1) Determine the number of cells for injection (ie 5´106 ) to determine the number of plates thatwill require trypsinizing (usually a 100% confluent p

Protocol-for-Aortic-Ring-Assay

ProceduresCover a 48-well plate with Matrigel (100μl/well) of and incubate for 30 min at 37°C, 5% CO2.Sacrifice the1-2 month old mice/rats (WT/mutant

alamarBlue®-Cell-Viability-Assay-Protocol

实验概要Cell health can be  monitored by numerous methods. Plasma membrane integrity, DNA  synthesis, DNA content, enzyme activity, presence of ATP, and c

RNA酶保护试验((RNase-Protection-Assay,RPA)简介

RNA酶保护试验((RNase Protection Assay,RPA)是通过液相杂交的方式,用反义RNA探针与样品杂交,以检测RNA表达的技术。1。原理:双链RNA(杂交的)能够抵抗RNA酶的降解。2。应用:检测RNA表达3。与Northern杂交和RT-PCR比较,RPA有以下几个优点:1.

RNA酶保护实验(RNase-Protection-Assay,RPA)简介

简介: RNA酶保护试验(RNase Protection Assay,RPA)是通过液相杂交的方式,用反义RNA探针与样品杂交,以检测RNA表达的技术。 1.  原理:双链RNA(杂交的)能够抵抗RNA酶的降解。 2.  应用:检测RNA表达 3.  与Northern杂交和RT-PCR比较

RNA酶保护试验((RNase-Protection-Assay,RPA)方法

一、试剂准备1. GACU POOL:取100mM ATP、CTP、GTP各2.78μl、100mM UTP 0.06μl,加DEPC H2O至100μl。2. 杂交缓冲液IPES 0.134g、0.5M EDTA(pH8.0)20μl、5M NaCl 0.8ml、甲酰胺8ml,加DEPC H2O至

RNA酶保护实验(RNase-Protection-Assay,RPA)简介

简介:RNA酶保护试验(RNase Protection Assay,RPA)是通过液相杂交的方式,用反义RNA探针与样品杂交,以检测RNA表达的技术。1. 原理:双链RNA(杂交的)能够抵抗RNA酶的降解。2. 应用:检测RNA表达3. 与Northern杂交和RT-PCR比较,RPA有以下几个优

James-Hardwicks-angiotensin-assay-protocol

 This specific procedure was developed to assay the activity of the Lck kinase expressed from a retroviral vector in rat fibroblasts. You can obviousl

RNA酶保护试验((RNase-Protection-Assay,RPA)的优缺点

RNA酶保护试验((RNase Protection Assay,RPA)是通过液相杂交的方式,用反义RNA探针与样品杂交,以检测RNA表达的技术。与Northern杂交和RT-PCR比较,RPA有以下几个优点:1. 检测灵敏度比Northern杂交高。由于Northern杂交步骤中转膜和洗膜都将造

RNA电泳实验方法

Polyacrylamide Gel Electrophoresis (PAGE)for use withRibonuclease Protection Assay (RPA):1. Making the Gel:  5% Denaturing gel for Ribonuclease Protec

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

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

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  

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

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

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.

Motility-Assay

DescriptionVarious phenotypic characteristics are requiredfor a cancer cell to successfully complete the metastaticcascade. Among these, acquisition o

Bradford-Assay

The bradford dye-binding assay is a colorimetric assay for measuring total protein concentration. It involves the binding of Coomassie Brilliant blue

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

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

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

Bradford-Assay

Bradford AssayThe bradford dye-binding assay is a colorimetric assay for measuring total protein concentration. It involves the binding of Coomassie B

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

Cytokine-Bioassays

Cytokine BioassaysIntroductionBiological activity of cytokines and their concentrations are commonly measured by cellular proliferation of primary cel

Neutralizing-Bioassay-Protocols

Neutralizing Bioassay ProtocolsIntroductionAntibodies that block binding of cytokines to their specific receptors and neutralize their effects are cri

NAi-protocol

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