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Unit Code 80178:
Scl 70 Antibodies, IgG, Serum

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Method Description

Recombinant Scl 70 antigen is bound to polystyrene microspheres,

which are impregnated with fluorescent dyes to create a unique

fluorescent signature. Scl 70 antibodies, if present in diluted serum,

bind to the Scl 70 antigen on the microspheres. The microspheres

are washed to remove extraneous serum proteins. Phycoerythrin

(PE)-conjugated antihuman IgG antibody is then added to detect

IgG anti-Scl 70 bound to the microspheres. The microspheres are

washed to remove unbound conjugate, and bound conjugate is

detected by laser photometry. A primary laser reveals the

fluorescent signature of each microsphere to distinguish it from

microspheres that are labeled with other antigens, and a secondary

laser reveals the level of PE fluorescence associated with each

microsphere. Results are calculated by comparing median

fluorescence response for Scl 70 microspheres to a 4-point

calibration curve. (Package insert:  Bioplex 2200 ANA Screen.

Bio-Rad Laboratories, Hercules, CA)

Performing Laboratory Location

Rochester

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