蛋白质技术

Materials 1.2 mM PIPESpH 7.0500 ml. 2.Gc-globulin (Calbiochem 345802)1 mg. 3.CFSE (Carboxyfluorescein succinimidyl ester; Molecular Probes). 4.Lysine100 mM in buffer 110 ml. 5.G25-150 desalting column ...

Considerations for use The Bradford assay is very fast and uses about the same amount of protein as the Lowry assay.It is fairly accurate and samples that are out of range can be retested within minut ...

Considerations for use The principle of the biuret assay is similar to that of the Lowryhowever it involves a single incubation of 20 min.There are very few interfering agents (ammonium salts being on ...

Considerations for use The bicinchoninic acid (BCA)assay is available in kit form from Pierce (RockfordIll.).This procedure is very applicable to microtiter plate methods.The BCA is used for the same ...

How long should cells be labeled? The ideal length of time to label cells depends on the protein of interest and the label that you are using.If you want to label an unstable protein with 35S-methioni ...

Recipes for the buffers are at the end of this protocol. 1.Label your protein with 32Pi.Then subject the protein to SDS polyacrylamide gel electrophoresis and transfer your gel-fractionated protein to ...

This specific procedure was developed to assay the activity of the Lck kinase expressed from a retroviral vector in rat fibroblasts.You can obviously use fewer or more cellsdepending on your needs.Don ...

Preparation of protein extracts for western blot 1.Grow cells to mid-log (OD600 less or equal to 1.0). 2.Harvest about 5 OD's of cells in a 13X100 glass dispo tube (Use new tubes.Can be up to about 6 ...

实验试剂： 试剂贮备液： 1、 1mol/L HCl48ml，Tris36.6g，TEMED0.23ml，加蒸馏水配成100ml，pH8.9。 2、 1mol/L HCl48ml，Tris5.98g，TEMED0.46ml，加蒸馏水配成100ml，pH6.7。 3、 Acr28g，Bis0.735g，加蒸馏水配成100ml。 4、 Acr10g，Bis2.5g，加蒸馏水配成100ml。 5、 核黄 ...

Definition Amino acids nucleotides polypeptides and other compounds in a colloidal state can be separated by the application of external voltages which cause charged colloid particles to move toward a ...

凝胶迁移或电泳迁移率实验（EMSA）是一种研究DNA结合蛋白和其相关的DNA结合序列相互作用的技术，可用于定性和定量分析。这一技术最初用于研究DNA结合蛋白，目前已用于研究RNA结合蛋白和特定的RNA序列的相互作用。通常将纯化的蛋白和细胞粗提液和32P同位素标记的DNA或RNA探针一同保温，在非变性的 ...

摘要：根据多肽固相合成的基本原理， 设计了2 套手工合成装置， 介绍了每套装置的操作方法， 并阐明了各种装置的优缺点和应用范围。

Q：SDS－PAGE电泳的基本原理？ A：SDS-聚丙烯酰胺凝胶电泳，是在聚丙烯酰胺凝胶系统中引进SDS（十二烷基硫酸钠），SDS能断裂分子内和分子间氢键，破坏蛋白质的二级和三级结构，强还原剂能使半胱氨酸之间的二硫键断裂，蛋白质在一定浓度的含有强还原剂的SDS溶液中，与SDS分子按比例结合，形成带负电荷的SDS-蛋白质复合物，这种复合物由于结合大量的SDS，使蛋白质丧失了原有的电荷状态形成仅保持原 ...

1. Rinse gel on glass plate briefly with distilled water. 2. Wash gel in fresh 50% ETOH-12% HAc with shaking 3 times 1 hour each. (Fix IEF gel ON in 10% TCA). 3. Wash gel four times with 10% ETOH-5% H ...

METHOD for Western Blots: 1.While your SDS-PAGE gel is runningmake your transfer buffer and chill to 4℃.Check that you have a frozen buffer dam is ready (stored in freezer next to Shikhatop shelf to t ...

DunnAnal.Biochem.1986: 157 GeorgiaTimes"1.5L GeorgiaTimes"1.0L GeorgiaTimes"10mM NaHCO3 ...

ECL or autoradiography? ECL is an appealing technique because it is quick and very sensitive and does not expose the investigator to radioactivity.The use of radioiodinated protein A to detect bound a ...

Materials •Suspension culture of fibroblast cells (1 liter) •35 mM Tris-HClpH 7.4140 mM NaCl (TBS buffer) •10 mM Tris-HClpH 7.510 mM KCland 1.5 mM magnesium acetate (TBS-M) •10X T ...

Motor proteins of several kinesin family groups have now been crystallized: monomeric Kinesin-1 motor domains from humanrat and Neurospora (Kull et al.1996; Sack et al.1997; Song et al.2001)and dimeri ...

Materials Milipore filter type HA 0.45 micron Culture plates (Linbro model 76-033-05) Protein in DDW or HEPES buffer (10 mg/ml) Vacuum grease Syringe with 18 G needle Procedure 1.Millipore filter a ...