Flow cytometric analysis of complement C4 binding protein (BP) on Group A Streptococcus pyogenes incubated with human C4 BP at a concentration of 10ug/ml for 1 hour at 37°C. Cells were harvested, washed with 1X PBS, and incubated with a complement C4 BP monoclonal antibody (Product # MA1-82815, black histogram) or isotype control (blue histogram) at a concentration of 2ug/ml for 1 hour at 37°C. For flow analysis, cells were washed in PBS, incubated with a fluorescently-conjugated donkey anti-mouse IgG secondary antibody at a dilution of 1:500 for 30 minutes at room temperature. Cells were washed with 1X PBS, fixed in 1% paraformaldehyde, and 10,000 cells were acquired for each sample. Data analysis was performed by gating on logarithmic fluorescence and side scatter (SSC) parameters. Data courtesy of the Innovators Program.
|Tested species reactivity||Human|
|Host / Isotype||Mouse / IgG2b|
|Immunogen||Native human C4 binding protein.|
|Storage buffer||BBS, pH 8.4|
|Contains||0.1% sodium azide|
|Storage Conditions||4°C or -20°C if preferred|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Flow Cytometry (Flow)||1-3ug/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
There are 3 variants of Complement component 4 binding protein (C4bp), which is composed of alpha and beta chains. The predominant form is a 570 kDa complex of 7 alpha chains and 1 beta chain. The other complexes formed are a 530 kDa homoheptamer of alpha chains or a 500 kDa complex of 6 alpha chains and 1 beta chain. C4bp regulates the activation of the classical complement pathway. It is a cofactor to the C3b/C4b inactivator, crucial to the proteolytic cleavage of C4b and important in cleavage of C3b. C4bp also regulates the C4bC2a complex (C3 convertase), accelerating its degradation by dissociating the complement fragment C2a. Both the C4bp alpha and beta chains interact with anticoagulant protein S and C4bp alpha binds protein S. C4bp is expressed in the blood serum.
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