By focusing on analysis techniques and protocols specific for Cancer Research, we are putting your research needs first. We will be featuring different cancer research tools throughout the year, providing you with the latest techniques and products.
Flow cytometry allows researchers to measure several physical characteristics of cells in suspension, such as cell shape, size, and internal complexity. By using a fluorescent compound to detect a specific cell component, the component can be further examined. For example, tumor cells often have abnormal amounts of DNA (aneupolid, i.e. having too many or too few chromosomes) compared to healthy diploid cells. This is the core application of flow cytometry in the area of Cancer Research.
The following functional assay is adapted from the book "Cancer Cell Culture: Methods and Protocols" ©2004 Humana Press, Product No. Z701424. This method is one of several detailed assays featured in this book. The book also contains detailed protocols on the isolation and culture of cancer cell lines.
Peter Mullen, Author
From Methods in Molecular Medicine, vol. 88 Cancer Cell Culture: Methods and Protocols, pp. 248-255
S. P. Langon, Editor
Humana Press Inc., Totowa NJ, Copyright 2004
Add 85.5 g sucrose, S7903, (250 mM) and 11.76 g trisodium citrate dihydrate, C5920, 2 H2O (40 mM) to a beaker of distilled water with 50 mLof dimethyl sulfoxide (DMSO), D2650. After adjusting to pH 7.6, fill to 1 L and store at 4 °C. This buffer is used to store frozen samples prior to analysis 1.
Dissolve 2 g trisodium citrate dihydrate, C5920, (3.4 mM), 2 mLIgepal® CA-630, I3021, (0.1% v/v), 1044 mg spermine tetrahyrdrochloride, S2876, (1.5 mM) and 121 mg Sigma 7-9, T1378, (0.5 mM) in 2 L of distilled water after adjusting to pH 7.6. This "Stock Solution" is used to make up the digestion/staining buffers.2
15 mg Trypsin, T0303, in 500 mLof Stock Solution (adjusted to pH 7.6).
250 mg Trypsin inhibitor, T9253, and 50 mg RNase A, R4875, in 500 mLof Stock Solution (adjusted to pH 7.6).
208 mg Propidium iodide, 81845, and 580 mg spermine tetrahydrochloride, S1141, made up in 500 mLof Stock Solution, adjusted to pH 7.6. Protect solution from the light by using metal foil throughout the preparation, storage, and the staining procedures (Note 2).
Igepal is a registered trademark of GAF Corporation.
FACSCalibur is a trademark of Becton Dickinson.
ModFit LT is a trademark of Verity Software House, Inc.
To continue reading please sign in or create an account.Don't Have An Account?