Detects mouse Complement Component C5a in direct ELISAs and Western blots. In direct ELISAs and Western blots (non-reducing conditions), approximately 50% cross-reactivity with recombinant human Complement Component C5a is observed.
Measured by its ability to neutralize Complement Component C5a-induced chemotaxis in the J774A.1 mouse reticulum cell sarcoma macrophage cell line [Zwirner, J. et al. (1998) Eur. J. Immunol. 28:1570]. The Neutralization Dose (ND50) is typically 0.6-3.0 µg/mL in the presence of 0.1 µg/mL Recombinant Mouse Complement Component C5a.
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Chemotaxis Induced by Complement Component C5a and Neutralization by Mouse Complement Component C5a Antibody. Recombinant Mouse Complement Component C5a (Catalog # 2150-C5) chemoattracts the J774A.1 mouse reticulum cell sarcoma macrophage cell line in a dose-dependent manner (orange line). The amount of cells that migrated through to the lower chemotaxis chamber was measured by Resazurin (Catalog # AR002). Chemotaxis elicited by Recombinant Mouse Complement Component C5a (0.1 µg/mL) is neutralized (green line) by increasing concentrations of Mouse Complement Component C5a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2150). The ND50 is typically 0.6-3.0 µg/mL.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Complement Component C5a
Mouse Complement 5a (C5a) is an enzymatically generated glycoprotein that belongs to a family of structurally and functionally related proteins known as Anaphylatoxins. C5a is a 77 amino acid (aa) peptide that is created by the activity of C5a convertase on the C5 alpha -chain (1, 2). Mouse C5a has four alpha -helices, plus three intrachain disulfide bonds that create a triple loop structure (3). In serum, proteolytic processing removes the C-terminal arginine, creating a low activity C5a desArg77 molecule (1). Mouse C5a shares 60% and 82% aa sequence identity to human and rat C5a, respectively. C5a binds to a signaling G-protein coupled receptor (GPCR) (C5aR/CD88), inducing neutrophil chemotaxis and endothelial cell activation (1, 4). It also triggers an oxidative burst in macrophages and neutrophils, and induces release of histamine in basophils and mast cells (1, 4). Alternatively, it may also bind to a non-signaling GPCR termed C5L2 whose function is yet to be determined (5).
Gerard, C. and N.P. Gerard (1994) Annu. Rev. Immunol. 12:775.
DiScipio, R.G. et al. (1983) J. Biol. Chem. 258:10629.
Huber-Lang, M.S. et al. (2003) J. Immunol. 170:6115.
Gerard, N.P. and C. Gerard (2002) Curr. Opin. Immunol. 14:705.
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