蛋白组学

Modified from that of Jay Thelen - University of Missouri-ColumbiaPhenol extraction followed by methanolic ammonium acetate precipitation - an effective protocol for sample preparation from protein-po ...

David M. SchieltzandMichael P. Washburn This protocol was adapted from "The Use of Mass Spectrometry in Proteomics" Chapter 8 inProteins and Proteomics(ed. Simpson). Cold Spring Harbor Laboratory ...

Lightning-Link 30 seconds hands on Very Easy to use 100% Antibody Recovery No separation steps Little antibody needed Easy to Scale Up Over 40 available labels http://www.youtube.com/v/w0XmU9Niwns& ...

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Sample PreparationFor Protein Concentration Determination of Cell Culture Decant medium from 10cm dish of adherent cells and rinse plate rapidly with phosphate-buffered saline (PBS). Aspirate excess P ...

Put the membrane in PBS or TBS with 7ul/ml B-mercaptoetanol and 2%SDS for 30' at RT in agitation Wash the membrane for 30' in PBS or TBS.Put the membrane in blocking buffer ...

Western Blotting with Biotinylated AntibodiesSAMPLE PREPARATIONFor Protein Concentration Determination of Cell CultureDecant medium from 10cm dish of adherent cells and rinse plate rapidly with phosph ...

Running Protein GelsSolutions10X Running Buffer (0.25 M Tris 1.92 M glycine 1% SDS)121 g Tris577 g glycine40 g SDSddh20 to 4 L (check pH at 1:10 dilution (pH=~8.8)).5X Sample Buffer (0.3125 M Tris pH ...

1. Run samples out on a gel. For bacterially expressed proteins generally 5 uL is plenty (1ml cell culture; cells resuspeded in 50 ul loading buffer). Run the gels (BioRad mini gels) at 195 V for appr ...

1. Run samples out on a gel. For bacterially expressed proteins generally 5 uL is plenty (1ml cell culture; cells resuspeded in 50 ul loading buffer). Run the gels (BioRad mini gels) at 195 V for appr ...

ANALYSIS OF PROTEINS BY IMMUNOPRECIPITATION P.J. Hansen Dept. of Animal Sciences University of Florida INTRODUCTIONImmunoprecipitation is a procedure by which peptides or proteins that react specifica ...

Antibody Precipitation with Protein A or GAuthor: James TurmoSource: Contributed by James TurmoAbstract: Using Immobilized Protein A or Protein G for Immunoprecipitation.PurposeUsing Immobilized Prote ...

CO-IMMUNOPRECIPITATION ANALYSIS OF PROTEIN-PROTEIN INTERACTIONSThis protocol uses total cell extracts to analyze putative protein-protein interactions in eukaryotic cells. One can also use nuclear ex ...

CO-IMMUNOPRECIPITATION ANALYSIS OF PROTEIN-PROTEIN INTERACTIONSThis protocol uses total cell extracts to analyze putative protein-protein interactions in eukaryotic cells. One can also use nuclear ex ...

Immunoprecipitation and Immune Complex kinase assay--according to Tamara Hurley 1) Wash cells with cold "Tris" (from Margie's shelf).Spin down the cells if you are working with suspension cells in a ...

Immunoprecipitation and Immune Complex kinase assay--according to Tamara Hurley 1) Wash cells with cold "Tris" (from Margie's shelf).Spin down the cells if you are working with suspension cells in a ...

Immunoprecipitation of metabolically labeled cells(Protocol from An Zhou Johns Hopkins 1995) 1. To your sample add Super E protease inhibitors 1 mM cold Met antibody. TMT extract or medium - dilute ...

Immunoprecipitation of metabolically labeled cells(Protocol from An Zhou Johns Hopkins 1995) 1. To your sample add Super E protease inhibitors 1 mM cold Met antibody. TMT extract or medium - dilute ...

Preparation of Modifed Radioimmunoprecipitation (RIPA) BufferRIPA Base IngredientsTris-HCl (buffering agent prevents protein denaturation) NaCl (salt prevents non-specific protein aggregation) NP-40 ( ...