Anti-CD59 Antibody, clone 2/24, FITC conjugated

BIOCHEMISTRY:

p I = 6.1

Human erythrocytes

Anti-CD59 Antibody, clone 2/24, FITC conjugated detects level of CD59 & has been published & validated for use in FC & IF.

Research Category
Inflammation & Immunology

Research Sub Category
Immunoglobulins & Immunology

This monoclonal is useful in studies of complement and homologous restriction of complement action of the properties of GPI linked proteins and in the characterisation of cells from patients with PNH.

SUGGESTED USAGE:
Flow cytometry and immunofluorescence – use 10µL direct from the vial per 1x10⁶ cells in a volume of 100 µL.
For analysis of erythrocytes, whole blood samples should be diluted approximately 500 fold.
For nucleated cells, use of the buffer coat will remove the burden of erythrocytes.
Platelets may be harvested by low speed centrifugation.

This antibody binds to a 17-25kD glycoprotein (Fletcher et al., 1990a,b) which is distributed on human cells and has a glycosyl-phosphatidylinositol (GPI) linkage to the plasma membrane(Hadam, 1989). It is identical to other CD59 monoclonals, MEM-43 and YTH 53.1, defined at the 4th International Conference on Human Leucocyte Differentiation Antigens(Davies et al., 1989; Stefanova et al., 1989). CD59 is an inhibitor of attack by autologous complement(Ojcius et al., 1990) possibly by limiting incorporation of C9 into the membrane complex(Rollins & Sims, 1990; Meri et al., 1990). GPI linked proteins such as CD59 are deficient on cells from patients with the acquired clonal disorder paroxysmal nocturnal haemoglobulinuria (PNH) in which the erythrocytes are susceptible to lysis by complement and leading to anemia(Rosse, 1990). Evidence suggests that CD59 may be the most important complement regulatory protein missing in PNH(Fletcher et al., 1990; Yamashina et al., 1990).

Cell reactivity (Fletcher et al., 1990a,b; Hadam, 1989):

The antibody reacts with all leucocytes, platelets, fibroblasts, erythrocytes and most cell lines. It is negative on U937 and Daudi cell lines. It is reduced on affected cells from patients with PNH with a distribution reflecting the clonal nature of the disease.

Purified immunoglobulin. FITC Conjugated. The antibody is supplied in phosphate buffered saline, pH 7.4, containing 0.2% bovine serum albumin and 0.1% sodium azide. The characteristics of each lot are tested by electrophoresis and flow cytometry.

STORAGE

Store at 2 to 8oC, for up to 6 months. For prolonged periods, store below -20oC in undiluted aliquots. AVOID REPEATED FREEZE/THAW CYCLES.

WARNING

The monoclonal reagent solution contains 0.1% sodium azide as a preservative. Due to potential hazards arising from the build up of this material in pipes, spent reagent should be disposed of with liberal volumes of water.

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