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染色质免疫沉淀法（Chromatin immunoprecitation，ChIP）是研究体内DNA与蛋白质相互作用的重要工具。它可以灵敏地检测目标蛋白与特异DNA片段的结合情况，还可以用来研究组蛋白与基因表达的关系。核小体组蛋白可以发生多种翻译后的共价修饰，如乙酰化、甲基化、磷酸化、泛素化等，这些共价修饰与真核基因的表达密切相关。根据“组蛋白密码”假说，组蛋白的各种共价修 ...

无论是从一个胚胎细胞分化为不同的组织器官，或者是从正常的组织突变为肿瘤组织，都可能涉及在同一基因组背景下不同基因的差异性表达。寻找差异表达的基因就有可能揭示细胞分化的机制或者肿瘤的成因，因而也就成为一项热门的技术。 寻找差异表达的基因有不少方法，抑制消减杂交（SSH）是比较有效的一种方法。以Clontech的PCR-Select cDNA Subtraction Kit为例：将样品（tester） ...

General Principles of Immunoprecipitation Lysis buffer. The choice of lysis buffer depends on what kind of IP you are doing. RIPA buffer gives the lowest backgroundbut can denature some kinases.It als ...

1)In the cold roomwash cells with cold "Tris" (the same stuff used for TC). Spin down the cells if you are working with suspension cells in a low speed swinging bucket centrifugeresuspend in ...

Lowry Protein Assay Laboratory of P.J.HansenDept.of Animal SciencesUniversity of Florida http://www.animal.ufl.edu/hansen/Protocols/LOWRY.htm The Lowry procedure is one of the most venerable and wide ...

Assemble the following components: •Cellsgrown in 10 cm dishes •Room Temp.PBS •Cold PBS •Cold Sonication Buffer (1% SDS10 mM EDTA50 mM Tris-HCl pH 7.5) Prepare sonication buffe ...

Purification of GST fusion proteins in E.coli GST融合蛋白纯化--纯化小量蛋白 Sugden labMcArdle Laboratory for Cancer ResearchUniversity of Wisconsin-Madison Medical School Small scale fusion protein preparation Gr ...

Buffer A Buffer B ...

主要特点： · 准确灵敏：BCA试剂的蛋白质测定范围是20－2000μg/ml采用加强方法可检测到5μg/ml；MicroBCA试剂测定范围是0.5-20μg/ml。 · 快速：45分钟内完成测定，比经典的Lowry法快4倍而且更加方便。 · 经济实用：除试管外，测定可在微孔板中进行，大大节约样品和试剂用量。 · 不受样品 ...

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 the following protocol for their own research obje ...

J. Movius K Coachman and S. Hahn (Hahn Lab) Induction of BRF in bacteria and purification on Ni-NTA agarose Transform BRF plasmid into strain BL21 DE3 (pLysS) or JM25 (DE3 containing the Arg t ...

Place the tissues on labeled aluminum foil and immediately place in dry ice.It is imperative that the tissues stay cold so that protease do not have time to act on the protein. Place the tissues in a ...

PIC Cross-linking and Immune Precipitation James FishburnHahn Lab March 2006 References Fishburn et.al.2005Molecular Cellvol.18 #3: Experimental Procedures pg.376 Immobilized Template assay (Hahn lab ...

Sample Buffer: 10 ml 0.06M Tris-HCl pH 6.8 0.6 ml 1M Tris 6.8 ...

1.Grow 25mls yeast cells to 5x10E6. 2.Sequentially spin down cells in a 15 ml polypro tube. 3.Wash cells with 1ml ice cold ddH2Otransferrring to an eppy tube. 4.Recon.cells in 1 ml ice cold ddH2O cont ...

HelveticaGenevaSwissSunSans-Regular" size="2"Linda Warfield Hahn Lab 2003 HelveticaGenevaSwissSunSans-Regular" size="2"Volumes given are for 2L of cells for each subunit (Toa1 and Toa2). Toa1 a ...

Purpose and backgrounds About nuclide... 35-S Decay: (b-ray0.167 MeV) Half life: 87.5 days Thick acrylic shield can block most of the emission.Because of the low energyit is impossible to detect spill ...

HIS-tagged protein purification John Mundy Institute of Molecular Biology Copenhagen Denmark http://www.arabidopsis.org/info/Protocols_Mundy2.jsp#his 1 liter cell prep 1) grow 20ml ...

Hancock Laboratory Methods.Department of Microbiology and Immunology University of British ColumbiaBritish ColumbiaCanada http://www.cmdr.ubc.ca/bobh/showmethod.php?methodid=28 MATERIALS: Running buff ...

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