MetabolicLabelingofCellswith35S

1) Transfer to a 24 wells plate the desired colonies.2) Once the cells are attached (at least 8 hours after tripsinizing them) add ~1 ml ofDME (met-, cys-).3) Add 3l of Met 35S, Cys 35S trans label.4) Incubate at least 5 hours at 370C (Put plate(s) inside a tupperware, containing abeaker with activated charcoal, and close loosely).5) Collect the media into 1.7 mL tubes and add 5l of the desired antibody.6) Incubate a......阅读全文

Metabolic-Labeling-of-Cells-with-35S

1) Transfer to a 24 wells plate the desired colonies.2) Once the cells are attached (at least 8 hours after tripsinizing them) add ~1 ml ofDME (met-,

Immunofluorescence-Labeling-of-Cells

实验概要Antibodies are an  important tool for demonstrating both the presence and the subcellular  localization of an antigen. Cell staining is a very ver

Sphingomyelin-Quantitation-Postcholine-Labeling-of-HL60-Cells

Lipid Extraction1) Following the appropriate time of treatment, transfer 4.5 ml into each of two duplicate glass pyrex tubes and maintain on ice.2) Sp

[3H]-Choline-Labeling-and-TNF-Treatment-of-HL60-Cells

1) Grow cells to a density of 5-8 X 105 cells/ml in RPMI 1640 containing serum.2) Pellet cells and wash 1 time with room temperature PBS.3) Resuspend

Labeling-Tubulin-and-Quantifying-Labeling-Stoichiometry

Labeling Tubulin and Quantifying Labeling StoichiometryThis is a general procedure for coupling moieties with reactive succinimidyl esters to tubulin.

Labeling-Tubulin-and-Quantifying-Labeling-Stoichiometry2

II. Labeling ProtocolThe procedure described below can be scaled down if desired. It is essential to perform all steps involving caged dyes under a sa

TUNEL-labeling

In Situ Cell Death (Apoptosis) Detection by TUNEL labelingby Boehringer Mannheim (Catalog No. 1684809), modified by Josiah N. Wilcox andJosé C. Rodrig

Biosynthetic-labeling

How long should cells be labeled? The ideal length of time to label cells depends on the protein of interest and the label that you are using. If you

Autoradiography-(35S)

Remove Kodak NTB2 nuclear emulsion from fridge and place at 42oC for around 30-60 mins (until melted).Make up the developer and the fixer and place in

Guide-to-Cell-Proliferation-and-Apoptosis-Methods

Chapter 1: Cell Death - Apoptosis and Necrosis1.1Introduction21.1.1Terminology of cell death21.1.2Differences between necrosis and apoptosis31.1.3Apop

Visceral-Fat-Deposits-and-the-Metabolic-Syndrome

Obesity is associated with many adverse health effects, including an increased risk of diabetes and heart disease. The combined condition of obesity,

Detection-by-TUNEL-labeling

In Situ Cell Death (Apoptosis) Detection by TUNEL labelingby Boehringer Mannheim (Catalog No. 1684809), modified by Josiah N. Wilcox,José C. Rodriguez

CMFDA-Labeling-of-Platelet

OUTLINECMFDA (5-chloromethylfluorescein diacetate) is a lipophilic tracer that has an enormous advantage over ordinary tracers (e.g. FITC) because it

Arachidonic-Acid-Labeling

1) Grow cells to a density of 5-8 X 105 cells/ml in RPMI 1640 containing serum.2) Pellet cells and wash 1 time with room temperature PBS.3) Resuspend

BrdU-Labeling-Protocol

实验概要The thymidine analog, 5-bromo-2-deoxyuridine (BrdU),is a common reagent used for cell proliferation assays and for the detection of apoptotic

In-vitro-Assessment-of-Metabolic--in-Suspension-Cryopreserved-Hepatocytes

实验概要BackgroundThe  pharmaceutical and biotechnology industry’s goal is to discover  therapeutic agents that are both safe and effective at treating or

Standard-Protocols-Autoradiography-(35S)

Remove Kodak NTB2 nuclear emulsion from fridge and place at 42oC for around 30-60 mins (until melted).Make up the developer and the fixer and place in

DNA-labeling-by-nick-translation

DNA labeling by nick translationreagents: DNA for labeling (concentration c > 150 ng/µl) modified nucleotides: Biotin-16-dUTP, Digoxigenin-11-dUTP, co

Basic-Method-for-Indirect-Immunofluorescence-Labeling

Basic Method for Indirect Immunofluorescence LabelingBackgroundThis is the method for indirect immunofluorescence labeling; that is, the antibodies do

南京大学化学化工学院谢然课题组诚聘化学生物学博士后

  课题组简介  谢然博士先后在上海交通大学董常明教授实验室(06-10),北京大学陈兴教授实验室(10-15),美国哈佛大学化学与化学生物学Daniel Kahne教授(美国两院院士)实验室(15-19)接受科研训练。主要研究方向包括糖生物学,蛋白质化学,生物正交反应、微生物学等。研究工作发表于S

Global-urinary-metabolic-profiling-procedures-using-gas-chromatography

Global urinary metabolic profiling procedures using gas chromatography–mass spectrometryEric Chun Yong Chan,1 Kishore Kumar Pasikanti1, 2 & Jeremy K N

Preparation-of-nucleic-acid-probes

Preparation of nucleic acid probesIn standard nucleic acid hybridization assays the probe is labeled in some way. Nucleic acid probes may be made as s

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

Differentiate-ES-cells-into-glial-cells-and-neurons

Day -1: Pass ES cells at normal density on gelatinized plate to free the culture of contamination fibroblast cells.___________________Day 1: Trypsiniz

ThiolReactive-Probe-Labeling-Protocol

实验概要Invitrogen  offers several fluorescent and biotinylated phalloidin and phallacidin  derivatives for labeling F-actin. These phallotoxins, isolated

Differentiating-Neural-Stem-Cells-into-Neurons-and-Glial-Cells

实验概要The protocols in  this section describe the steps involved in differentiating neural stem  cells (NSC) to neurons, astrocytes, and oligodendrocyte

线粒体荧光探针大全:TMRM,Mitotracker,JC1(1)

线粒体荧光探针信息大全 (Probes for Mitochondria)包括各种常用探针,如JC-1,JC-9,TMRM,TMRE等Mitochondria are found in eukaryotic cells, where they make up as much as 10% of th

Culture-of-Endometrial-Explants

Culture of Endometrial Explants and Peri-implantation Conceptuses to Monitor Synthesis and Secretion of Proteins and ProstaglandinsP. J. Hansen and J.

Growing-cells

No two cell lines behave exactly the same, so you must learn the peculiarities, or personality, of each of the cell lines with which you work. Irrespe

Lyophilizing-Cells

Inoculate 200 ml L-broth supplemented with appropriate antibiotics with the bacteria to be lyophilized.Incubate the culture at 37°C with vigorous shak