Figure. The binding activity of C1qB with PFN1. The complement component 1q (or simply C1q) is a protein complex involved in the complement system, which is part of the innate immune system. C1q together with C1r and C1s form the C1 complex. Antibodies of the adaptive immune system can bind antigen, forming an antigen-antibody complex. When C1q binds antigen-antibody complexes, the C1 complex becomes activated. Activation of the C1 complex intitiates the classical complement pathway of the complement system. The antibodies IgM and all IgG subclasses except IgG4 are able to initiate the complement system. C1q is composed of 18 polypeptide chains: six A-chains, six B-chains, and six C-chains. Complement Component 1, Q Subcomponent B (C1qB) is six B-chains of C1q. Besides, Profilin 1 (PFN1) has been identified as an interactor of C1qB, thus a binding ELISA assay was conducted to detect the interaction of recombinant human C1qB and recombinant human PFN1. Briefly, C1qB were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100μL were then transferred to PFN1-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-C1qB pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50µL stop solution to the wells and read at 450nm immediately. The binding activity of C1qB and PFN1 was shown in Figure 1, and this effect was in a dose dependent manner.
用法
Reconstitute in 20mM Tris, 150mM NaCl (pH8.0) to a concentration of 0.1-1.0 mg/mL. Do not vortex.