摘要:1.Fix gel in 50% methanol for greater than 1 hour rinse gel in dH2O 3 times 5 minutes each place back in 50% methanol for 1 hour.2.Stain gel in 100 mls of stain solution for 30 minutes 3.Rinse gel in [阅读全文:]
摘要: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 needleProcedure1.Millipore filter all solu[阅读全文:]
摘要:Materials •Suspension culture of fibroblast cells (1 liter)•35 mM Tris-HCl,pH 7.4,140 mM NaCl (TBS buffer)•10 mM Tris-HCl,pH 7.5,10 mM KCl,and 1.5 mM magnesium acetate (TBS-M)•10X [阅读全文:]
摘要: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 an[阅读全文:]
摘要:1.While your SDS-PAGE gel is running,make your transfer buffer and chill to 4℃.Check that you have a frozen buffer dam is ready (stored in freezer next to Shikha,top shelf to the right side--looks lik[阅读全文:]
摘要:Dunn,Anal.Biochem.1986: 157 1.5L 1.0L10mM NaHCO31.26gchill to 4oC0.84g3mM Na2CO30.477gpH ~9.90.318gin 20% MeOH300mL200mL1.Blot: no need to presoak gell.Transblot: 1-2H @ 1A keep T<35℃Miniblot: 1H @ 25[阅读全文:]
摘要: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% [阅读全文:]
摘要: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[阅读全文:]
摘要:The purpose of this method is to separate proteins according to their size, and no other physical feature. In order to understand how this works, we have to understand the two halves of the name: SDS [阅读全文:]
摘要:凝胶迁移实验以下问题是以Promega公司试剂盒为例。1)什么是凝胶迁移或电泳迁移率实验？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[阅读全文:]