Product #: K002

CH50 Test

Complement hemolytic activity is a functional test of the classical and alternative pathway of complement in plasma or serum.

CH50 test kit (100 determinations)

European Union Price: €520,- (As per 01-01-2024).
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CH50 Test
Description

Complement haemolytic activity is a functional test of the classical and alternative pathway of complement in plasma or serum. The classical pathway method (CH50) is based on lysis of sensibilized sheep erythrocytes in the presence of Ca++ and Mg++. It is helpful as screening test when complement depletion or deficiency is suspected. The test is sensitive to the reduction, absence and/or inactivity of any component of the pathway.

This method is also suited to assess the effects of pharmaceuticals on inhibition or consumption of complement components. Additionally, haemocompatibility of biomaterials and medical devices according to the international standard ISO 10993-4:2017 after blood, plasma, or serum contact with biomaterials can be evaluated. Activation of the complement system by biomaterials or pharmaceuticals can result in consumption of complement proteins, which reduces the CH50 level.

Application

Measuring classical pathway complement activity. Plasma or serum samples from the following species can be analyzed: human, guinea pig, hamster, rat, rabbit, bovine, swine, M. rhesus and beagle. This kit is intended for laboratory research use only and is not for use in diagnostic or therapeutic procedures.

Principle

An erythrocyte suspension is incubated for 30 minutes with serial diluted serum or plasma at 37 ºC. The activation of the complement system will result in haemolysis. After incubation the samples are centrifuged to obtain supernatant. The free haemoglobin concentration in the supernatant, which is directly proportional to complement activity, is measured by means of a spectrophotometer at a wavelength of 415 nm. Plotting the dilution factor of plasma against the degree of haemolysis allows the calculation of the CH50 (i.e. the dilution of serum to obtain 50% lysis of erythrocytes).

The positive reference is total lysis induced by lysis fluid and the negative reference is obtained after incubation with dilution buffer.

The kit is designed to determine complement activity via the classical pathway of small samples (25 µL) and can be performed in a 96 well (round bottom) microplate. The assay can also be performed in microcentrifuge tubes.

Video protocol for the CH50 Test kit.
Storage and Stability

Product is stable at 4 °C for at least six weeks.

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Manual / SDS

References that used the CH50 assay:
  1. Wu WK, Ukita R, Patel YJ, Cortelli M, Trinh VQ, Ziogas IA, Francois SA, Mentz M, Cardwell NL, Talackine JR, Grogan WM, Stokes JW, Lee YA, Kim J, Alexopoulos SP, Bacchetta M. Xenogeneic cross-circulation for physiological support and recovery of ex vivo human livers. Hepatology. 2023 Sep 1;78(3):820-834.

  2. Sülzen H, Began J, Dhillon A, Kereïche S, Pompach P, Votrubova J, Zahedifard F, Šubrtova A, Šafner M, Hubalek M, Thompson M, Zoltner M, Zoll S. Cryo-EM structures of Trypanosoma brucei gambiense ISG65 with human complement C3 and C3b and their roles in alternative pathway restriction. Nat Commun. 2023 Apr 27;14(1):2403.

  3. Lang Balija M, Štimac A, Košutić Gulija T, Gudan Kurilj A, Bekavac A, Plećaš A, Halassy B, Jagušić M, Forčić D. Evaluation of the Interactions between Mumps Virus and Guinea Pig. J Virol. 2023 Apr 27;97(4):e0035923.

  4. Wu WK, Stier MT, Stokes JW, Ukita R, Patel YJ, Cortelli M, Landstreet SR, Talackine JR, Cardwell NL, Simonds EM, Mentz M, Lowe C, Benson C, Demarest CT, Alexopoulos SP, Shaver CM, Bacchetta M. Immune characterization of a xenogeneic human lung cross-circulation support system. Sci Adv. 2023 Mar 31;9(13):eade7647.

  5. Weisser NE, Sanches M, Escobar-Cabrera E, O’Toole J, Whalen E, Chan PWY, Wickman G, Abraham L, Choi K, Harbourne B, Samiotakis A, Rojas AH, Volkers G, Wong J, Atkinson CE, Baardsnes J, Worrall LJ, Browman D, Smith EE, Baichoo P, Cheng CW, Guedia J, Kang S, Mukhopadhyay A, Newhook L, Ohrn A, Raghunatha P, Zago-Schmitt M, Schrag JD, Smith J, Zwierzchowski P, Scurll JM, Fung V, Black S, Strynadka NCJ, Gold MR, Presta LG, Ng G, Dixit S. An anti-HER2 biparatopic antibody that induces unique HER2 clustering and complement-dependent cytotoxicity. Nat Commun. 2023 Mar 13;14(1):1394.

  6. Khosousi S, Hye A, Velayudhan L, Bloth B, Tsitsi P, Markaki I, Svenningsson P. Complement system changes in blood in Parkinson’s disease and progressive Supranuclear Palsy/Corticobasal Syndrome. Parkinsonism Relat Disord. 2023 Mar;108:105313.

  7. Quint WH, Matečko-Burmann I, Schilcher I, Löffler T, Schöll M, Burmann BM, Vogels T. Bispecific Tau Antibodies with Additional Binding to C1q or Alpha-Synuclein. J Alzheimers Dis. 2021;80(2):813-829.

  8. Pinnapireddy SR , Giselbrecht J , Strehlow B , Janich C , Husteden C , Meister A , Loppnow H , Sedding D , Erdmann F , Hause G , Brezesinski G , Groth T , Langner A , Bakowsky U , Wölk C . A triple chain polycationic peptide-mimicking amphiphile - efficient DNA-transfer without co-lipids. Biomater Sci. 2019 Dec 17;8(1):232-249.

  9. Giselbrecht J, Wiedemann S, Reddy Pinnapireddy S, Goergen N, Loppnow H, Sedding D, Erdmann F, Bakowsky U, Hause G, Lúcio M, Langner A, Wölk C. Nucleic acid carrier composed of a branched fatty acid lysine conjugate-Interaction studies with blood components. Colloids Surf B Biointerfaces. 2019 Dec 1;184:110547.

  10. Garaulet G, Lazcano JJ, Alarcón H, de Frutos S, Martínez-Torrecuadrada JL, Rodríguez A. Display of the Albumin-Binding Domain in the Envelope Improves Lentiviral Vector Bioavailability. Hum Gene Ther Methods. 2017 Dec;28(6):340-351.

  11. Akhouri RR, Goel S, Furusho H, Skoglund U, Wahlgren M. Architecture of Human IgM in Complex with P. falciparum Erythrocyte Membrane Protein 1. Cell Rep. 2016 Feb 2;14(4):723-736.

  12. Misra SK, Chang HH, Mukherjee P, Tiwari S, Ohoka A, Pan D. Regulating Biocompatibility of Carbon Spheres via Defined Nanoscale Chemistry and a Careful Selection of Surface Functionalities. Sci Rep. 2015 Oct 14;5:14986.

  13. Mukherjee P, Misra SK, Gryka MC, Chang HH, Tiwari S, Wilson WL, Scott JW, Bhargava R, Pan D. Tunable Luminescent Carbon Nanospheres with Well-Defined Nanoscale Chemistry for Synchronized Imaging and Therapy. Small. 2015 Sep;11(36):4691-703.

  14. Luo J, Lindstrom J. Antigen-specific immunotherapeutic vaccine for experimental autoimmune myasthenia gravis. J Immunol. 2014 Nov 15;193(10):5044-55.