CellThawing／CellFreezingProtocol
Freezing Cells:Cells should be growing well or known to be in log phaseCount, collect and pellet cells in a 15mL test tubeResuspend in freezing media so that the concentration is no more than 5x10^6 cells/mL of cold freezing mediaTransfer 1mL of cells to appropriately labelled cryovials and maintain on ice for approximately 30minutesTransfer vials to -80C freezer for 24hrsTransfer to liquid nitrogen dewar or -140C fr......阅读全文
Nucleolar－Isolation－Protocol
We recommend that you first download and read this page as a PDF file. Using that as your guide, you can then follow the protocol below and view a Qui
Sandwich－ELISA－Protocol
实验概要The Sandwich ELISA measures the amount of antigen between two layers of antibodies (i.e. capture and detection antibody). The antigen to be mea
Dot－blot－protocol
实验概要A technique for detecting, analyzing, and identifying proteins, similar to the western blot technique but differing in that protein samples are
Yale－Immunofluorescence－Protocol
实验概要We provide a protocol for fixation, immunostaining, and imaging in 384-well Plates.主要试剂Reagents1. 384-well view plates (Aurora)2. HUVEC (pooled, L
Immunofluorescence－Microscopy－Protocol
实验概要Immunofluorescence allows the imaging of a specific factor in cells or tissue sections through the use of a specific antibody chemically which i
Immunofluorescence－Microscopy－Protocol
实验概要Immunofluorescence allows the imaging of a specific factor in cells or tissue sections through the use of a specific antibody chemically which i
Histone－blotting－protocol
实验概要 Western blot detection of histone proteins. 实验步骤 The following protocol refers to the western blot detection of histone proteins derived from p
Urea－Lysis－Protocol
Urea lysis buffer 9M Urea, 2.5mM EDTA, 2.5mM EGTA, 1% DTE, 4% CHAPS make 10ml and aliquot 10x1ml, freeze at -70°C Lysate prepara
RNA－Isolation－Protocol
RNA Isolation Protocol(Revised 5-15-2003)Stabilize RNAStart with 15 ml E. coli Culture containing 7.5* 109 cells (OD600= 0.2 Dilute cells or scale up)
Cytotoxicity－Assays－Protocol
Cytotoxicity Assays ProtocolCell-mediated cytotoxicity was determined by using a standard microcytotoxicity assay. Briefly, target cells were pelleted
Intracellular－Staining－Protocol
1. Fix cells- Add16% formaldehyde directly into culture medium to obtain a final concentration of 1.5% formaldehyde.2. Incubate in fixative for 10 min
Basic－ELISA－Protocol
实验概要 There are many different types of ELISAs, which can detect the presence of protein in serum or supernatent. One of the most common typ
Adhesion－Assay－Protocol
Materials to be prepared beforehand:1) Washing Buffer--0.1% BSA in medium (DMEM or RPMI)2) Blocking Buffer--0.5% BSA in medium (DMEM or RPMI)3) Lamini
Bacteria－Culture－Protocol
Bacteria Culture ProtocolBy 徐晓政1、TBS Medium Preparation:Prepare 1L of TBS medium contains:Tryptone 12gYeast extract 24gNaCl 5gSodium Succinate 5gGlyce
Colony－PCR－Protocol
1. Pull out eight glycerol stock plates from the –80oC freezer and set on bench top to thaw. Be sure to remove the foil seal before leaving the plates
Silver－Staining－Protocol
1x 40min - overnight 50% MeOH, 12% Acetic Acid1x 30min 50% MeOH, 12% Acetic Acid, 0.05% 37% Formaldehyde3x 20min
TAIL－PCR－Protocol
TAIL is a series of reactions that are intended to map where a T-DNA (transfer DNA) has inserted within the genome. The main components of the 3 react
Midiprep－preparation－of－Plasmid－DNA
实验概要The PureLink™ HiPure Plasmid DNA Midiprep Kit allows purification of 100–350 μg of high-quality plasmid DNA from 15–25 mL overnight E. coli cul
The－Impact－of－Harmo...－（一）
实验概要During more than 25 years of application in immunological sciences, ELISPOT has been established as a routine, robust, versatile, and reliable a
Growth,－Maintenance－and－Transfection－of－Suspension－Adapted－293EBNA－cells
ProcedureI. INTRODUCTIONThe 293 EBNA cell line is established from primary embryonal human kidney cells transformed with sheared human adenovirus type
Maxiprep－preparation－of－Plasmid－DNA
实验概要The PureLink™ HiPure Plasmid DNA Maxiprep Kit allows purification of 500–850 μg of high-quality plasmid DNA from 100–200 mL overnight E. coli cu
Culture－of－BEND－Cells－(Bovine－Endometrial－Cells)
Culture of BEND Cells (Bovine Endometrial Cells)Charles E. Krininger, III and Peter J. Hansen Dept. of Animal Sciences, University of FloridaThis prot
CellTrace™－CFSE－Cell－Proliferation－Kit
实验概要The CellTrace™ CFSE Cell Proliferation Kit provides a versatile and well-retained cell-tracing reagent in a convenient and easy-to-use form. The
TOP10－chemically－competent－cells
OverviewThis protocol is a variant of the Hanahan protocol [1] using CCMB80 buffer for DH10B, TOP10 and MachI strains. It builds on Example 2 of the B
Tissue－Culture－Methods1
I. TYPES OF CELLS GROWN IN CULTURETissue culture is often a generic term that refers to both organ culture and cell culture and the terms are often us
Xenograft－Tumor－Assay－Protocol
1) Determine the number of cells for injection (ie 5´106 ) to determine the number of plates thatwill require trypsinizing (usually a 100% confluent p
Simplified－Arabidopsis－Transformation－Protocol
(Brief version for those who are familiar with the method)Steve Clough and Andrew Bent, University of Illinois at Urbana-Champaign.Our present proto
Cajal－Body－Isolation－Protocol
Buffers and solutions(All solutions are supplemented with Complete Protease inhibitor tablet (Roche, Cat no: 1-873-580) at the final concentration of
Tissue－preparation－protocol－for－ChIP
实验概要This protocol describes how chromatin is prepared from tissue, which can subsequently be used for chromatin immunoprecipitation (ChIP). It is re
Marcantonio－Lab－Protocol－Manual
Protein GelPreparing and Running a Protein Gel (7% Polyacrylamide) A. Preparation of Running Gel Solution 1) Add to a 50 ml cylinder:DD-H2O 26 ml30% a