Baby Rabbit Complement, Standard Grade

Multiple freeze-thaw cycles will result in reduced complement potency, so we offer a variety of pack sizes to satisfy applications ranging from small scale assay development experiments to large scale clinical studies. We recommend limiting the number of freeze-thaw cycles and sub-aliquot into single-use quantities whenever possible in order to extend the shelf life of all of our complement products.

We offer baby rabbit and human complement in both forms. Frozen liquid complement is ready to use after thawing and is stored at -70°C or below. Lyophilized complement is stored at -20°C or below, is easier to ship, and is reconstituted with sterile water to the label volume, or to a smaller volume when you need higher potency.

Yes. Incubate the cells with a complement-fixing antibody to the lineage you want removed (anti-Thy-1.2, anti-CD4 or anti-CD8 for mouse T cells, for example), add baby rabbit complement at about 1:10, hold 1 hour at 37°C and wash away the lysed cells; one round typically removes over 95% of the target population. Complement from 3-4-week-old rabbits has low heterophile antibody activity, so the untreated cells are spared. Titrate the antibody and the complement on your cells first.

The hemolytic titer measures how well complement lyses antibody-sensitized sheep red blood cells; CH50 is the dilution of serum that lyses 50% of the cells. It is the release test for every lot of our complement products, and our heat-inactivated human products are CH50 tested to confirm that complement activity is absent.

Work backwards from the readout. Complement regulators are host-specific, so the complement source is part of the assay method, not a preference.

• Antibody-depleted human complement for the hSBA (serogroup B meningococcus and gonococcus; correlate hSBA ≥ 1:4). Neisseria recruits human factor H, which rabbit and guinea pig factor H cannot replace, and the donor antibodies in native human serum would kill the target on their own.
• Baby rabbit complement for the rSBA (serogroups A, C, W and Y; correlate rSBA ≥ 1:8) and for pneumococcal OPA/MOPA, which was standardized on it.
• Adult rabbit complement for complement-dependent cytotoxicity and HLA crossmatching, the classical cell-lysis source.
• Guinea pig complement where potency per volume is the point: CH50 titration, complement fixation and the vibriocidal assay.

When the model animal's own cascade is the readout, use matched-species complement. Mouse, rat, pig, canine, cynomolgus, goat and hamster complement are available on request through our custom quote form.

For what the research literature actually uses, by pathogen and assay, see which complement source for which assay.

Endogenous complement activity can cause intrinsic, non-specific bacterial killing. We recommend preventing this by heat-inactivating all OPA/OPKA antisera at 56°C for 30 minutes, and always running controls of complement, HL60 cells and bacteria without antiserum to assess non-specific killing. If you are still seeing higher than expected non-specific killing, try the following:
• Run the assay with a new aliquot of bacteria, in case the original was compromised in a way that made it easier to kill.
• Use a different bacterial strain to see if you can replicate the finding.
• Test a new HL60 cell preparation, preferably at a higher or lower passage number.