ProtocolforCellFusion
Healthy Sp2/0 cells should be rapidly growing by this time. Sp 2/0 cells should be started about two weeks before the cell fusion. Every two days, they should be centrifuged at a 64.4 xg on the IEC clinical centrifuge (in 50 ml tubes) for two minutes. The media should be removed and the cells resuspended in fresh media. This ensures that only the healthiest cells will fall to the bottom of the tube. Centrifugation at......阅读全文
The－OP9DL1－System:－Generation－of－TLymphocytes－from－Embryonic2
Freezing ESCs 25. Passage the ESCs as described in Step 22. Resuspend the cells in 3 mL of freezing medium for OP9 cells. 26. Aliquot 1 mL of cell sus
Cell－cycle－analysis－of－Escherichia－coli－cells
Cell cycle analysis of Escherichia coli cellsC period = the time for a round of chromosome replicationD period = the time between the end of a round o
Cellbased－ELISA－for－primary－cells
1. The 96-well micro-plates are pre-coated with extra cellular matrix.2. Primary cells are grown in the 96-well plates. Seed 100μl of 10,000-20,000
LIVE/DEAD®－Violet－Viability/Vitality－Kit
实验概要The LIVE/DEAD® Violet Viability/Vitality Kit provides a two-color fluorescence cell viability and vitality assay that is based on the simultaneo
Gram－Stain－(+\)
实验概要细菌的革兰氏染色技术实验材料Colonies of bacteriaToothpicksCrystal violetGram's iodine95% ethanolSafraninMicroscopes with oil immersion实验步骤1. Before staini
Human－Embryonic－Stem－(ES)－Cell－Protocols——General－notes－on－ES－cell－culture
hES media has a two week shelf life.hES cells should be cultured in 4, 6, 24, 48, 96 well plates. Growing cells in flasks is not recommended because i
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
TCell－Activation－Using－mAb－to－CD3
IntroductionMature T cells recognize and respond to the antigen/MHC complex through their antigen-specific receptors (TCR). The most immediate consequ
Culture－of－Human－Embryonic－Stem－Cells－(hESC)
All cell lines are initially grown according to the supplier's protocols but we are adapting them to one simple protocol outlined below:6-well pla
Application－Note:－Qdot®－Nanocrystal－Conjugates－in－Flow－Cytometry
实验概要Researchers today are trying to maximize the information that they get out of flow cytometry experiments by looking at more parameters in a sing
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
SQ－Blood－Mini－Protocol－for－50ul－Clotted－Blood
实验概要This protocol is designed for isolating genomic DNA from 0.5-1 million cultured cells. For larger or smaller amounts of starting cell numbers,
Bacteria,－cont.－Growth－and－Reproduction
Flagella and motilitymonotrichous flagella - the bacterial cell has a single flagellaperitrichous flagella - the bacterial cell has several flagella w
Bacteria－Growth－and－Culture－Bacteria－Growth－and－Culture
Flagella and motilitymonotrichous flagella - the bacterial cell has a single flagellaperitrichous flagella - the bacterial cell has several flagella w
Intracellular－Cytokine－Staining－Protocol
实验概要A modification of the basic immunofluorescence staining and flow cytometric analysis protocol can be used for the simultaneous analysis of surf
CELL－CYCLE－ANALYSIS
PROPIDIUM IODIDE: The most commonly used dye for DNA content/cell cycle analysis is PROPIDIUM IODIDE (PI). It can be used to stain whole cells or isol
病毒冷冻保藏技术
实验概要Snap freezing, or flash freezing, is the process by which samples are lowered to temperatures below -70°C very rapidly using dry ice or liquid
Detection－of－Intracellular－Antigens－by－Flow－Cytometry
实验概要Fix and Perm reagents are designed for use with all commercially available flow cytometers. Alignment and compensation should be performed accor
Vybrant®－DyeCycle™－Ruby－stain
实验概要Live cell studies of cellular DNA content and cell cycle distribution are useful to detect variations of growth patterns due to a variety of phy
Human－FastTrack－mRNA－Isolation
Preparation of Cells1.Prepare or collect between 2x107 cells for each mRNA prep (will yield about 10-20µg of mRNA). If PBMCs from a whole blood sample
Growing－Overnight－Cultures
1. Place 2 mL of the appropriate sterile medium in a 13 mm yellow-capped culture tube. If more culture is needed, place up to 5 mL in a 16 mm green-ca
酵母转化的几种方法
Modified Yeast Transformation Inoculate cells from an overnight culture into 50 ml YEPD and incubate at 30°C with shaking. Typically, add 0.1 to 0.2 m
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
Western－blotting样品准备
实验概要Preparation of lysis buffers, protease and phosphatase inhibitors, lysate from cell culture, lysate from tissues, protein concentration, samples
Bacterial－transformation
IntroductionTransformation is the process of introducing foreign DNA (e.g plasmids, BAC) into a bacterium. Bacterial cells into which foreign DNA can
LselectinPNAd－Interactions－under－Flow－Conditions.
PurposeThe main purpose of the flow chamber assay is to visualize and measure interactions between flowing cells expressing a given adhesion molecule
Flow－Cytometry－of－Fibroblast－Nuclei－for－DNA－content
MaterialsP.I. Solution: 4 mM Na3Citrate (0.118 g/100 mL)30 U/mL RNAseI (43 mg/100 mL)0.1% Triton-X100 (0.1mL/100 mL)50 µg/mL propidium iodide (5 mg/10
Method:－Reactivating－Cell－Lines－and－Cell－Growth－for－DNA－Preparation
Purpose:Cell lines are reactivated and grown to a count of 1 x 108 cells. The cells are pelleted and stored frozen at -80 degrees C prior to DNA extra
E.Z.N.A.®－Protocol－for－Bacteria
实验概要The E.Z.N.A. Total RNA Kit can be modified for isolation of RNA from bacterial cultures. Only cells growing at log phase should be used. Measu
Stem－cell－characteristics－of－amniotic－epithelial－(AE)－cells
Isolation of AE Cells1. Human placentae were obtained with the approval of the institutional review board, after uncomplicated elective caesarean