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MaterialsRIPA buffer (RIPA buffer enables the extraction of cytoplasmic, membrane and nuclear proteins and is compatible with many applications, including reporter assays, protein assays, immunoassays and protein purification. RIPA Buffer does not contain protease or phosphatase inhibitors. However, if desired, protease and phosphatase inhibitors can be added to the RIPA buffer just before use to prevent proteolysis ......閱讀全文
This page should point you to the many different general and lab-specific protocols describing tissue and cell lysis and serve as forum for comparison
Fine Mapping of Genomic Targets of Nuclear Proteins in Cultured CellsAchim Breiling and Valerio OrlandoDulbecco Telethon Institute, Institute of Genet
實驗概要Native chromatin immunoprecipitation to query specific chromatin states of individual genes. 主要試劑10 x TBS 0.1 M Tris-HCl (pH 7.5) 1.5 M NaCl
實驗概要The method is a native chromatin immunoprecipitation protocol.主要試劑1. 10 x TBS 0.1 M Tris-HCl (pH 7.5) 1.5 M NaCl 30 mM CaCl2 20 mM MgCl2 50 mM Na
1. The preparation of native chromatin from cultured human cells1.1.Cultured cells (e.g. HL-60 or lymphoblastoids) are grown to a density of approxima
實驗概要The PureLink? Genomic DNA Purification Kit allows rapid and efficient purification of genomic DNA. The kit is designed to efficiently
實驗概要Preparation of lysis buffers, protease and phosphatase inhibitors, lysate from cell culture, lysate from tissues, protein concentratio
IntroductionIn our lab we use the term 'cell-free system' when we talk about the examination of apoptotic activity in cytoplasmic extrac
1) After ECL development, wash membrane once for 10min with PBST.2) Incubate the membrane in stripping buffer (see below) in a heat-sealed plastic bag
實驗概要 This protocol utilizes the powerful guanidine isothiocyanate–phenol:chloroform extraction method which allows the
實驗概要Interest in RNA-protein interactions is booming as we begin to appreciate the role of RNA, not just in well-established processes such as tran
實驗概要We provide a general IP procedure including a list of reagents and a table to help you choose the correct protein beads.Immunoprecipit
My Experience Purifying RNA from E. coliRegarding RNA extraction, there is a horrible tendency of people to use kits for RNA extraction with bacteria.
實驗概要 The immunoprecipitation (IP) of cross-linked chromatin with antibodies specific for certain histone modifications (chromatin &nb
實驗概要ChIP is a powerful tool that allows the specific identification of proteins or histone modifications to regions of the genome. Chromat
Cellular Transfection and Immunoprecipitation Before proceeding with the experiments outlined below, all kinase pocket mutants should be ch
Induction of BRF in bacteria and purification on Ni-NTA agaroseTransform BRF plasmid into strain BL21 DE3 (pLysS) or JM25 (DE3 containing the Arg tRNA
實驗概要E.Z.N.A.? Total RNA Midiprep Kit provides a rapid and easy method for the isolation of up to 600 ug of total RNA from cultured eukaryo
III. Methods for DNA isolationA. Large scale double-stranded DNA isolationThe method used for the isolation of large scale cosmid and plasmid DNA is a
實驗概要Preparation of lysis buffers, protease and phosphatase inhibitors, lysate from cell culture, lysate from tissues, protein concentratio
實驗概要The E.Z.N.A.? Blood DNA Maxi Kit is designed for isolation of genomic DNA from up to 25 ml of fresh, whole blood treated with any comm
Phosphorylation of SubstratesScott T. Eblen, N. Vinay Kumar, and Michael J. WeberDepartment of Microbiology and Cancer Center, University of Virginia
實驗概要Ideally, one would like to be able to directly phosphorylate substrates in an intact cell. This could potentially be performed by intr
Preparation of Glutathione-S-Transferase (GST) Fusion ProteinsMargret B. Einarson and Elena N. Pugacheva Foxx Chase Cancer Center, Philadelphia,
RNA提取（主要內容如下）Tips for Handing RNA Total RNA IsolationmRNA IsolationrRNA IsolationOthersQ & A posted in the Method Forum Basic Proc
Introduction The most important aspect of our cloning vectors is that t
實驗概要The E.Z.N.A.? Total RNA Maxi Kit uses HiBind? matrix spin-column technology to isolate up to 5 mg total cellular RNA from a variety of
實驗概要The Dynabeads? and buffers provided in this kit will enable you to a) covalently immobilize antibodies of your choice onto the surface
實驗概要The Dynabeads? and buffers provided in this kit will enable you to a) covalently immobilize antibodies of your choice onto the surface
Metabolic biotinylation of mammalian cell receptors for imagingBakhos A. Tannous , btannous@hms.harvard.edu, Massachusetts General Hospital and Harvar