Monoclonal Antibodies and Conjugates to Human Leukocyte Markers

Sigma Monoclonal Antibodies to Human Leukocyte Markers (CD Antigens) are evaluated for their performance in flow cytometry and/or immunohistochemical staining. Unless otherwise indicated, the products are supplied as purified mouse immuno globulins in 0.01 M phosphate buffered saline, pH 7.4, containing 1% BSA and 0.1% sodium azide as preservative.

Human CD Antibodies and their conjugates are accompanied by a product data sheet providing detailed protocols for immunofluorescence staining and flow cytometry applications.

The CD designation defines the antigen cluster. These CD Antibodies have been evaluated for their specificity by the 2nd, 3rd, 4th and/or 5th International Workshop on Human Leucocyte Differentiation Antigens except as noted. Summaries of these data have been published and can be found as follows:

1.  Leucocyte Typing II, Reinherz, E.L., et al., (eds.), Springer Verlag, New York (1986).

2.  Leucocyte Typing III, McMichael, A.J., et al., (eds.), Oxford University Press, Oxford (1987).

3.  Leucocyte Typing IV, Knapp, W., et al., (eds.), Oxford University Press, Oxford (1989).

4.  Leucocyte Typing V, Schlossman, S.F., et al., (eds.), Oxford University Press, Oxford (1995).

CD Antibody - Conjugates

Sigma Biotin conjugated CD antibodies are prepared by conjugation of biotinamidocaproate N-hydroxysuccinimide ester with purified mouse immunoglobulin fraction from ascites fluid. Biotin conjugated antibodies are evaluated in flow cytometry. The products are supplied in 0.01 M phosphate buffered saline, pH 7.4, containing 1% BSA and 0.1% sodium azide as preservative.

Sigma FITC conjugated CD antibodies are prepared by conjugation of fluorescein isothiocyanate isomer I (FITC) with the purified mouse immunoglobulin fraction from ascites fluid. The conjugates are purified by gel filtration and contain no detectable free FITC. The F/P molar ratio of the conjugates is 3.0 - 8.0. FITC- conjugated CD antibodies are evaluated in flow cytometry. The products are supplied in 0.01 M phosphate buffered saline, pH 7.4, containing 1% BSA and 0.1% sodium azide as preservative.

Sigma R-Phycoerythrin (PE) conjugated CD antibodies are prepared by conjugation of R-phycoerythrin with the purified mouse immunoglobulin fraction from ascites fluid, according to a modification of the method of Kronick.(1) PE-conjugated CD antibodies are evaluated in flow cytometry. The products are supplied in 0.01 M phosphate buffered saline, pH 7.4, containing 1% BSA and 0.1% sodium azide aspreservative.

Quantum Red^TM is a tandem fluorescent dye created by coupling R-phycoerythrin with Cy5 reactive dye. This dye is efficiently excited at 488nm and emits at 670nm. It is useful as a third color in combination with fluorescein and

R-phycoerythrin for multiparameter analysis. It is useful as a second color in combination with fluorescein when minimal compensation requirement between the green and red photomultiplier tubes is desired. Quantum Red is also useful as the single color when there is high autofluorescence of the cells. Sigma Quantum Red conjugated CD antibodies are prepared by conjugation of Quantum Red to purified mouse immunoglobulin from ascites fluid, according to a modification of the method of Kronick.(1) Quantum Red-conjugated antibodies are evaluated by flow cytometry and formulated to obtain maximum percent positive staining without significant elevation of background staining. The products are supplied in 0.01 M phosphate buffered saline, pH 7.4 containing 1% BSA and 0.1% sodium azide as preservative.

Reference:

1.  Kronick, M.N., J. Immunol. Meth., 92, 1 (1986).

U.S. Patent No. 4,859,582; 4,542,104

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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the Purdue University Cytometry Laboratories and distributed free of charge as an educational service to the cytometry community. If you have any comments please direct them to Dr. J. Paul Robinson, Professor & Director, PUCL, Purdue University, West Lafayette, IN 47907. Phone:(765) 494-0757; FAX (765) 494-0517; Web http://www.cyto.purdue.edu, EMAIL cdrom3@flowcyt.cyto.purdue.edu