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MICROVUETMBa EIA

The MICROVUE Ba Enzyme Immunoassay Kit measures the amount of the complement fragment Ba in human urine, plasma or serum.

Product Specifications

Background

Measurement of Ba in human urine, plasma or serum provides evidence of the involvement of the alternative pathway of complement. The alternative complement pathway provides innate protection against microbial agents in the absence of specific antibody. The activation of this complement pathway can be triggered by a variety of substances including microbial polysaccharides or lipids, gram-negative bacterial lipopolysaccharides, and surface determinants present on some viruses, parasites, virally infected mammalian cells, and cancer cells. In autoimmune diseases, the alternative complement pathway may contribute directly to tissue damage. A centrally important reaction that occurs during alternative pathway activation is the conversion of the 93 Kd molecular weight Factor B zymogen to an active proteolytic enzyme. This is accomplished in a two-step reaction. During the first reaction step the Factor B forms a magnesium-dependent complex with C3 (H20) or C3b. The C3 (H20), B complex is formed only in fluid-phase while the C3b, B complex can be formed either in fluid-phase or on a target surface. Factor B, which is present in the C3 (H20), B or the C3b, B complex, is cleaved into the Ba (33 Kd) and Bb (60 Kd) fragments in the second reaction step by the alternative pathway enzyme, Factor D. Although alternative pathway activation is thought to occur primarily in the absence of specific antibody, many situations arise in which alternative pathway activation can occur as the result of classical pathway activation. For example, immune complexes that are present in autoimmune disease patients can trigger classical complement pathway activation with resultant production of C3b fragments. As described above, these C3b molecules are capable of binding Factor B and initiating its cleavage into the Ba and Bb fragments. Thus, alternative pathway activation can occur in antibody-mediated autoimmune disease states and may contribute significantly to enhanced complement activation and concomitant tissue destruction. By assessing Factor B cleavage products in test specimens, one can estimate the extent of alternative pathway utilization occurring at the time of sample collection in the disease state under investigation. The MICROVUE Ba EIA provides a simple, rapid, non-radioactive, highly specific, and quantitative procedure for measuring Factor B activation. It is ideal for investigations involving the role or status of the alternative complement pathway in numerous research and clinical settings and for monitoring the generation of Ba in vitro.

Cross Reactivity

African Green Monkey, Cynomolgous monkey, Dog, Rhesus monkey

Assay Performance Time

2.5 hours

Standard

5

Sample Type

Serum 10 μL, EDTA Plasma 25 μL, Urine 10 μL

Sample Volume

Serum 436–3362 ng/mL, EDTA Plasma 226–2153 ng/mL, Urine 0.6–27.0 ng/mL

Limit Of Detection

0.011 ng/mL

Form

96 well plate with 12 eight-well strips in a resealable foil pouch

Storage Conditions

Store the unopened kit at 2°C to 8°C. Refer to Product Insert for additional storage details.

Limit Of Quantification

3.239 ng/mL

Controls

2

Inter Assay

2.4–8.1%

Intra Assay

2.2–3.3%

Frequently Asked Questions

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