SphingomyelinQuantitationPostcholineLabelingofHL60Cells
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) Spin down cells in the table top centrifuge at 1,200 rpm, 40C, for 5 minutes.3) Gently aspirate and discard the spent media (remember, consider it radioactive). Be extremely careful not to disturb the pellet since it is somewhat dispersed.4) Immediately upon asp......阅读全文
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
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
AA－－Metabolite－Quantitation－of－Media－PostAA－Labeling
1) Remove 2 500 µl aliquots of supernatant into scintillation vials, add scintillation fluid and count.2) Aliquot 1.6 mls of the remaining supernatant
AA－－Metabolite－Quantitation－of－Cell－Pellets－PostAA－Labeling
Extraction:1) Following spin, save supernatant for analysis. Be extremely careful not to disturb the pellet since it is somewhat dispersed.2) Immediat
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-,
[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.
Purification－of－Demethylated－Sphingomyelin
I. Lower, chloroform phase:1) Dry on rotovapor system with house vacuum lines. It is not necessary to dry sample completely, but sufficiently to yield
Sphingomyelin－Mass－Measurement
Protocol:Bligh & Dyer extraction1) Pellet approximately 1 X 107 cells.2) Resuspend pellet in 3 ml CHCl3: CH3OH (1:2) and vortex hard.3) Add 0.8 ml H2O
Labeled－Sphingomyelin－Synthesis
Synthesis reaction:1) Mix purified DMSM with cyclohexylamine and 14CH3I at a ratio of 1/1.1/1.3 in 5 ml of methanol.2) Allow reaction to proceed at ro
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
Column－Purification－of－Demethylated－Sphingomyelin
Packing column:1) To 20 g of 100-200 mesh Bio-Sil A silica gel add 80 mls of chloroform.2) Place a small portion of glass wool at the base of the colu
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
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
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
Multiple－studies－with－a－single－experiment:－The－Power－of－...（四）
Better Ion Transmission With Segmented Quadrupole Segmented Quadrupole •Improved transmission across m/z range and for narrow windows•Brighter Source
Multiple－studies－with－a－single－experiment:－The－Power－of－－...（一）
Multiple studies with a single experiment: The Power of Quantitative MultiplexingMultiple studies with a single experiment: The Power of Quantitative
寡核苷酸的相关操作
In this section, you will find techniques related to oligonucleotides, such as oligo purification by acrylamide gel, annealing two oligos to make doub
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
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
P2RY8基因编码功能及结构描述
该基因编码的蛋白属于g蛋白偶联受体家族，它优先被腺苷和尿苷核苷酸激活。该基因在未分化的HL60细胞中中度表达，位于X染色体和Y染色体上。The protein encoded by this gene belongs to the family of G-protein coupled recept
Multiple－studies－with－a－single－experiment:－The－Power－of－...（五）
Competitive Advantages Trust your quantitation! Multinotch MS3 quantitation is more accurate than other MS2 Methods The accuracy of Multinotch MS3 qua
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
P2RY8基因突变与药物因子介绍
该基因编码的蛋白属于g蛋白偶联受体家族，它优先被腺苷和尿苷核苷酸激活。该基因在未分化的HL60细胞中中度表达，位于X染色体和Y染色体上。[由RefSeq提供，2008年7月]The protein encoded by this gene belongs to the family of G-pro