AA&MetaboliteQuantitationofCellPelletsPostAALabeling
Extraction:1) Following spin, save supernatant for analysis. Be extremely careful not to disturb the pellet since it is somewhat dispersed.2) Immediately upon removing supernatant from each tube, add 3 mls of chloroform-methanol, 1:2, and vortex vigorously!!--> It is necessary to add the C:M immediately in order to get good dispersion of the pellet. If too much time passes prior to resuspension, the pell......阅读全文
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
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
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
In－Situ－Cell－Death－(Apoptosis)－Detection－by－TUNEL－labeling
Protocol for Frozen Sections:Warm 150ml 4% Paraformaldehyde/1x PBS to RT. Fix slides in it, 20 min., RT.1x PBS rinse, 2 times.1x PBS, 30 min., RT. Beg
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.
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
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
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
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
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
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
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
A－convenient－method－for－the－isolation－of－crude－nuclear－pellets.
This procedure describes a convenient method for the isolation of crude nuclear pellets from N. crassa. The method, an adaptation of the one developed
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
体外荧光法检测核内体早期动力学5
Leave tubes on ice and repeat Steps 13–15 for the next 6–12 plates of cells.When all cells are collected, centrifuge all tubes at 250g for 5 min at 4
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
Basic－Method－for－Indirect－Immunofluorescence－Labeling
Basic Method for Indirect Immunofluorescence LabelingBackgroundThis is the method for indirect immunofluorescence labeling; that is, the antibodies do
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
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-,
Multiple－studies－with－a－single－experiment:－The－Power－of－...（六）
•SCX and high pH reversed phase fractionation are both orthogonal to low pH C18 LC separation•Strong cation exchange (SCX) requires sample desalting
活细胞荧光成像的新型标记法及其在STED中的应用（五）
SNAP-tag技术在STED超高分辨率显微成像中的应用近十年中，显微成像技术得到了飞跃的发展，填补光学显微镜（~200 nm）到电子显微镜（~0.1 nm）分辨率缺口，打破光学衍射极限的超高分辨率显微镜也越来越趋于成熟化。其中，德国马普研究所的Stefan Hell教授凭借其研发的受激发射
Protocol－for－Dual－Pulse－Labeling－Using－EdU－and－BrdU－Incorporation
实验概要The measurement of cell proliferation is fundamental to the assessment of cell health, genotoxicity, and drug efficacy. Proliferation is traditi
Isolation－of－Total－RNA－from－Animal－Cells－use－RNeasy－Mini－Kit
实验概要Extract the total RNA from animal cells by using RNeasy Mini Kit (QIAGEN No.74104) 主要试剂SDS based extraction solution实验步骤1. Harvest cells.1) Try
E.coli－Total－RNA－Labeling－Protocol－for－Spotted－Microarray
Note:Start with 20 ug of total RNA for each labeling reaction.All solutions that can be filtered should be filtered.Cy dyes are light sensitive and sh
Immunofluorescent－Staining－of－Mouse－and－Rat－Leukocytes
I. ProcedureHarvest cells from tissue, preparing a single cell suspension. Red blood cells may be removed by lysis or density gradient: Red blood cell
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