细胞功能测定

Fixation1) Collect 2 X 106 cells.2) Pellet cells by spinning at 1000 rpm 4°C for 5 minutes.3) Resuspend cell pellet in 1 ml of cold PBS.4) Fix cells by adding 4 ml of -20°C absolute ethanol.5) Store c ...

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/100 mL)ProcedureHarvest cells: Rinse with a subconfl ...

MATERIALS:1. 1 X PBS (PBSAz 1 X PBS e.g. Irvine Scientific CA containing 2% newborn calf serum and 0.1% sodium azide)2. 7-Amino-actinomycin D (7-AAD e.g. Calbiochem CA)3. Human AB serum heat-inactiv ...

Protocol Wash cells with PBS and trypsinize to a single cell suspension. Count an aliquot on a hemocytometer. Meanwhile centrifuge the cells (e.g. 1000 RPM 5 min.) to pellet them. Resuspend the cells ...

Mary-Ann Campbell's method. This is a quick method of getting paraformaldehyde (PFM) into solution. It is a good alternative to the traditional methods which can take hours and has the advantage that ...

DescriptionProcedure for CD3/ CD4 /CD8 T cell subset counts using CD3-FITC / CD4-PE / CD8-PECy5 antibody Procedure1. One 5 ml round bottom tube (Falcon) is taken per blood sample to be analyzed. The t ...

METHOD 1:Formaldehyde preservative � 2% formaldehyde solution in protein-free phosphate-buffered saline (PBS).Prepare as follows:Add 2 g paraformaldehyde powder (e.g. Sigma St. Louis MO) to 100 ml o ...

Use this Protocol for Directly or Indirectly staining cells for Flow Cytometric Analysis 1) Dilute cells to 5x10^6 cells/mL2) Aliquot 100uL of cells per per tube (5x10^5 cells total)3) Add 20uL blocki ...

ReagentsCells to be studied expressing green fluorescent protein (GFP). Note that the same cell type without GFP is needed as a control.1 X PBS2% Buffered formaldehyde solution (see recipe)70% Ethan ...

ReagentsCells to be studied expressing green fluorescent protein (GFP). Note that the same cell type without GFP is needed as control.Hoechst 33342 stock solution (1mg/ml) (see recipe)12 X 75 mm cul ...

DescriptionProtocol for intracytoplasmic staining of cytokines for FACS analysis Procedure1) Prepare spleen lymph node or T cell clone cells as single cell suspension using erythrocyte lysis (water) o ...

DescriptionCell Fixation staining and flow cytometric analysis ProcedureCells (106) were washed twice in FACS buffer (phosphate buffered saline PBS pH 7.4 containing 2% fetal calf serum(FCS) and 0.02% ...

DescriptionThis protocol is for use with the D cyclins and employs 488 nm argon laser excitation of propidium iodide and 630 nm NeNe or diode laser excitation of the fluorochrome Cy5 to detect cell cy ...

Wash cells with PBS Fix cells in 0.2% glutaraldehyde for 5’ dilute this in PBS Wash cell with PBS Stain with X-gal solution (ingredients listed below) for 6 to 24 hours at 37°C Chemical Conc.Stocks ...

DescriptionCell Staining for Senescence-Associated â-Galactosidase (SA-â-Gal) Activity Procedure1. Carefully remove the growth media from the cell culture. 2. Add an equal volume of PBS and gently mix ...

Materials Fibroblast cells in log phase growthCa Mg free-phosphate buffered saline (PBSA)5% (w/v) Glutaraldehyde (GTA)2% (w/v) Perchloric Acid (PCA)Subbed slides (coated with chrom alum gelatin) and P ...

before start:thaw cells from liquid nitrogen grow in 75cc flask (T75) in Fischer's medium MM (maintenance medium) until content gets cloudy and starts to turn yellow (usually want ~ 6 x 106 cells per ...

This is a simple assay useful for obtaining quantitative information about the relative density of cells adhering to multi-well cluster dishes. The dye in this assay crystal violet stains DNA. Upon so ...

a cell suspension is mixed with trypan blue and examined by low-power microscopy Materials cells PBS M3 hemocytometer 0.4 % trypan blue in PBS micropipet microscope Protocol prepare cell suspension at ...

DescriptionCytotoxic activities of mNK cells are examined in 51Cr release assay from target cells ProcedureEffector cells (mNK cells) are seeded into round-bottom 96-well plates (Costar) in 100-?l ali ...