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[General Lab] Alkaline Phosphatase And Phospholipase C Assays 日期：2005-07-18 00:00:00 点击：176 好评：0
Wednesday, October 01, 2003 Description Alkaline phosphatases (APs) are highly ubiquitous enzymes, present in all species from bacteria to man Procedure 1.Grow bacteria overnight in Pi sufficient and Pi deficient media. 2.Read OD600 of ove...
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[Enzymology] Detection of Phospho-Hsp27 in THP.1 cells by Bioplex Assay 日期：2005-07-18 00:00:00 点击：315 好评：0
Friday, June 11, 2004 Description Medium throughput assay for detection of the phosphorylation of Hsp27 by upstream kinases (notably MapKap Kinase 2) Procedure Phospho-HSP27 Cellular Assay Cell Type: THP-1 cells from ATCC (TIB-202) that ha...
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[Enzymology] A generic Phosphatase and pyrophosphatase assay 日期：2005-07-18 00:00:00 点击：250 好评：0
Sunday, December 28, 2003 Description This review discusses the EnzChek phosphatase assay kit comercially available from Molecular Probes Inc. that we have found to be very valuable to assay probable phosphatase activity of enzymes. The as...
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[Enzymology] Growth, Purification, and Refolding of&n 日期：2005-07-18 00:00:00 点击：283 好评：0
Description Growth, Purification, and Refolding of TGF-b3 (for 3L) Procedure 1) Transform BL21(DE3)cells with pET32 plasmid containing the TGF-b3 sequence a) Place 10ml competent cells in a microfuge tube and add ~0.5mg plasmid DNAon ice....
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[Enzymology] Procedure for Ion (DEAE) Exchange C 日期：2005-07-18 00:00:00 点击：284 好评：0
Description Procedure for Ion (DEAE) Exchange Chromatography for ecT bR2 Purification Procedure For cells from a 3.0 liter culture: 1. Prepare 400ml of 50mM Imidazole/0.6MNaCl/pH 7.0 2. Next, set up one 45 ml bed volumecolumn of DEAE-Sepha...
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[Enzymology] Buffer Exchange Procedure for the Preparation of NMR Samples 日期：2005-07-18 00:00:00 点击：318 好评：0
Friday, November 21, 2003 Description The preparation of a biomolecule for NMR studies usually requires exchanging the sample in an aqueous solution appropriate for conducting the proposed NMR studies. The two main requirements that must b...
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[Enzymology] Growth of Prototrophic E. coli Strains in D2O Minimal Salts 日期：2005-07-18 00:00:00 点击：270 好评：0
Friday, November 21, 2003 Description The following is a protocol for growing 1.0 L of E. coli in deuterium oxide (D2O)-M9 salts medium. Due to the slow growth of cells in D2O minimal medium, the D2O culture is inoculated with cells at 0.3...
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[Enzymology] Analysis of soluble/insoluble partitioning of E. coli overex 日期：2005-07-18 00:00:00 点击：263 好评：0
Friday, November 21, 2003 Description Analysis of soluble/insoluble partitioning of E. coli overexpressed proteins Procedure Pellet 1.0 mL of induced cell culture in a microfuge tube. Remove supernant. Resuspend the cells in 0.5 mL of MQ H...
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[Enzymology] Sephadex G-50 spun column purification 日期：2005-07-18 00:00:00 点击：347 好评：0
Wednesday, November 19, 2003 Description Spin column purification can be used to change buffers without a concomitant change in solution volume, to remove protein contaminants or to purify plasmid DNA suitable for sequencing. Procedure 1)...
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[Enzymology] Yeast Nuclear Extract 日期：2005-07-18 00:00:00 点击：179 好评：0
Tuesday, November 18, 2003 Description Yeast Nuclear Extract Procedure Cell Growth 3 liters of cells are grown in YPD (3% glucose) to A600 of 3-5. Antifoam A (two drops from a P200 pipetman) is added to media before autoclaving. For wild t...