Eccles:ProteinLysatesfromTissue

Cell Lysis Buffer5mL 0.1M Tris HCl pH 8 (10mM)0.44g NaCl (150mM)0.02g EDTA (1mM)0.5mL nonidet P40 (1% w/v)0.05g SDS (0.1% w/v)Make up to 50mL with MQH2O and filter sterilise.20x Complete mini Protease InhibitorTake 1 tablet and dissolve in 0.5mL MQH2O by pipetting up and down.Store on ice.Dilute 20 fold in cell lysis bufferCell Lysis Buffer containing 1x Complete Protease Inhibitor50uL 20x Complete protease inhibitor......阅读全文

Eccles:Protein-Lysates-from-Tissue

Cell Lysis Buffer5mL 0.1M Tris HCl pH 8 (10mM)0.44g NaCl (150mM)0.02g EDTA (1mM)0.5mL nonidet P40 (1% w/v)0.05g SDS (0.1% w/v)Make up to 50mL with MQH

Eccles:Protein-Lysates-from-Cells-in-Culture

Cell Lysis Buffer 5mL 0.1M Tris HCl pH 8 (10mM) 0.44g NaCl (150mM) 0.02g EDTA (1mM) 0.5mL nonidet P40 (1% w/v) 0.05g SDS (0.1% w/v) Make up to 50mL

PCR-from-Tissue

collect piece of tissue (e.g., pieces of a leaf or flower) in Eppendorf tube containing 40 ml 0.25 N NaOH put in boiling H2 O for 30 sec (optimu

PCR-from-Tissue

collect piece of tissue (e.g., pieces of a leaf or flower) in Eppendorf tube containing 40 ml 0.25 N NaOHput in boiling H2O for 30 sec (optimum may ne

PCR-from-Tissue

1.collect piece of tissue (e.g., pieces of a leaf or flower) in Eppendorf tube containing 40 ml 0.25 N NaOH 2.put in boiling H2O for 30 sec (optimum

PCR-from-Plant-Tissue

1.protocol (1)collect piece of tissue (e.g., pieces of a leaf or flower) in Eppendorf tube containing 40 ml 0.25 N NaOH (2)put in boiling H2Ofor 30

PCR-from-Plant-Tissue

PCR from Tissue Reference:  Klimyuk et.al., 1993, Plant J. 3:493-494 Last updated: 1/27/00 By: Kay Schneitz        collect piece of tissue (e.g., piec

DNA-Extraction-from-Tissue

实验概要DNA extraction from tissue.主要试剂Extraction buffer100 mM Tris-HCl (pH 8.0)     100 mM EDTA (pH 8.0) 100 mM Na-Phosphate (pH 8.0)   1.5 M NaCl1% CTAB

Dissociation-of-Cells-from-Primary-Tissue

实验概要 A  common method to obtain single cell suspensions from primary tissue is  enzymatic disaggregation. Expose the cells to enzymes for a minimal

RNA-Collection--Purification-from-fibrous-tissue

实验概要 Provides an easy and fast method for isolating total RNA from fibrous tissues which contains contractile proteins, connective tissue and co

DNA-Extraction-from-Frozen-Tissue-Sections

Tissue collection, storage, microdissection, sectioning: See separate protocol.Tissue handling: Note that all fresh tissue should be handled as BioSaf

Cell-and-tissue-lysis-hub

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

RNA-Isolation-From-Animal-tissue-or-cell-culture

实验概要This method is  designed for most animal tissues and culture cells. For RNA isolation  from fibrous tissue, follow the specialized protocol on pag

Protein-extraction-from-whole-tissues-for-IEF

Modified from that of Jay Thelen - University of Missouri-ColumbiaPhenol extraction followed by methanolic ammonium acetate precipitation - an effecti

Isolation-of-Genomic-DNA-from-Tissue-Using-ChargeSwitch®-Technology

实验概要 The ChargeSwitch®  gDNA Mini and Micro Tissue Kits allow rapid and efficient purification  of genomic DNA from mini (10-25 mg) or micro (3-5 mg)

Isolation-of-stromal-vascular-cells-from-human-adipose-tissue

Stromal stem cells proliferate in vitro and may be differentiated along several lineages. The scarcity of stromal stem cells in tissues and the la

Signaling-Pathway-from-GProtein-Families

G-aS-coupled receptors stimulate adenylyl cyclase (AC), which synthesizes cAMP from ATP. In contrast Gai-coupled receptor

Cell-Extraction-Protocol

实验概要Primary tissues  are valuable tools for the study of intracellular and extracellular  markers which characterize disease states. We have developed

Immunoprecipitation...-(一)

实验概要We provide a  general IP procedure including a list of reagents and a table to help  you choose the correct protein beads.Immunoprecipitation is a

RNA-Purification-from-1020-mg-Paraffinembedded-Tissue

实验概要 The E.Z.N.A.®  SQ Tissue RNA Kit is designed for isolating total RNA from animal  tissue and cultured cells. The solution based system can be e

Chemical-Induction-of-Apoptosis

Chemical Induction of Apoptosis - 1 May 2001p53, p21WAF1, Myc, Bcl-2, Bax, Bcl-x and bak are among the proteins involved in the regulation of apoptosi

Dephosphorylation-...

实验概要 The method  provides a protocol for removal of phosphate groups from proteins,  before or after blotting. To demonstrate the specificity of an

Western-blotting样品准备-(一)

实验概要Preparation of  lysis buffers, protease and phosphatase inhibitors, lysate from cell  culture, lysate from tissues, protein concentration, samples

RNA-extraction-using-trizol/tri

RNA extraction with TRIzol (Invitrogen product name) or the equivalent TRI (Sigma-Aldrich product name) is a common method of total RNA extraction fro

Genomic-DNA-Extraction--PureLink™

实验概要The  PureLink™ Genomic DNA Purification Kit allows rapid and efficient  purification of genomic DNA. The kit is designed to efficiently isolate  g

同位素法测定底物磷酸化活性方法

实验概要Ideally, one would  like to be able to directly phosphorylate substrates in an intact cell.  This could potentially be performed by introducing AT

Yeast-Lysates-for-Westerns

Cells are grown for 2-3 days as 1.5ml prep. under selection for the plasmid of interest. Spin cells down 2.6K for 5min. Resuspend in 1ml 0.25m N

Embryo-Lysates--Immunoprecipitation

Embryo lysatesTake 25 embryos and place into 1.7ml centrifuge tube.Rinse once in lysis buffer (add ~ 1ml) and remove by aspirationAdd 500 µL lysis buf

Preparation-of-Phage-Lysates

Preparation of Phage LysatesInoculate 5 ml of lambda-broth in a glass culture tube with a single colony of an appropriate host strain of E. coli. Incu

同位素法测定底物磷酸化活性方法-Phosphorylation-of-Substrates

Phosphorylation of SubstratesScott T. Eblen, N. Vinay Kumar, and Michael J. WeberDepartment of Microbiology and Cancer Center, University of Virginia