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Active Complement Component 1, R Subcomponent (C1r)

C1-r; complement Component 1; Complement component 1 subcomponent r

ACTIVITY TEST

The classical complement pathway plays a major role in innate immunity against infection. This pathway is triggered by C1, a multimolecular complex composed of the recognition protein C1q and two serine proteases, C1r and C1s. Following the C1q recognition, C1r is autoactivated, and in turn activates C1s, which cleaves C4 and C2, the C1 substrates. Both C1r and C1s activation involve cleavage of a specific Arg-Ile bond, converting single-chain proenzymes into active proteases of disulfide bond-linked chains (A and B). The full-length (amino acid residues 1-707) of mouse C1r was expressed which activity was measured by its ability to cleaves a thioester substrate Z-Lys-SBzl•HCl. The reaction was performed in 50 mM Tris, pH 7.5 (Assay Buffer), initiated by addition 50 μL of various concentrations of C1r (diluted by Assay Buffer) to 50 µl substrate mixture of 0.2mM Z-Lys-SBzl•HCl and 0.2 mM DTNB. The final well serves as a negative control with no C1r, replaced with 50 μl assay buffer. Then read in kinetic mode for 5 minutes at an absorbance of 405 nm. The specific activity of recombinant mouse C1r is > 500 pmol/min/µg.

USAGE

Reconstitute in 10mM PBS (pH7.4) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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