Standard Curve
Example of Mouse Complement C5a standard curve in Assay Diluent TA2. Plotted are the background-subtracted data.
This standard curve is provided for demonstration only.
Mouse Complement C5a OneStep competitive ELISA Kit
Mouse Complement C5a OneStep competitive ELISA Kit
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Product Details
Product Details
Product Specification
| Immunogen | Recombinant Protein |
| Antibody Type | Recombinant mAb |
| Reactivity | Ms |
| Purification | Protein A |
| Stability & Storage | 12 months from date of receipt / reconstitution, 2 to 8°C as supplied. |
Kit
| Precision | Intra-assay: 3.7%； Inter-assay: 6.5% |
| Sample type | Serum; EDTA Plasma, Heparin Plasma, Citrate Plasma; Cell Culture Supernatant |
| Assay type | Sandwich (quantitative) |
| Sensitivity | 3.52 ng/mL |
| Range | 15.6 ng/mL – 1000 ng/mL |
| Recovery | Serum: 108% EDTA Plasma: 104% Heparin Plasma: 108% Citrate plasma: 86% Cell Culture Supernatant: 105% |
| Assay time | 60 minutes |
| Species reactivity | Ms |
| CROSS REACTIVITY | Rt, Gp |
Background
Mouse Complement C5a is a 77-amino acid peptide generated during complement activation through the cleavage of the α-chain of complement component C5. It is a potent anaphylatoxin and chemoattractant that belongs to a family of structurally related proteins involved in innate immunity. Mouse C5a exerts its effects primarily by binding to the G-protein coupled receptor C5aR (CD88), which is widely expressed on leukocytes and various other cell types. It induces neutrophil chemotaxis, endothelial cell activation, and triggers an oxidative burst in macrophages and neutrophils. Additionally, C5a stimulates histamine release from basophils and mast cells and mediates IL-8 release from bronchial epithelial cells. Mouse C5a shares 60% and 82% amino acid sequence identity with human and rat C5a, respectively. It also plays a role in hepatocyte regeneration and has been implicated in various inflammatory and immune-mediated diseases.
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ELISA
Spike Recovery
The recovery of Mouse Complement C5a was evaluated in activated samples spiked with concentrations spanning the entire assay range.
Dilution Linearity
The concentrations of Mouse Complement C5a were measured and interpolated from the target standard curves and corrected for sample dilution.
The leftmost bar in the graph shows biological samples diluted in Assay Diluent to 50% concentration. "undiluted" was defined as 50% biological sample. The subsequent bars represent two-fold serial dilutions of this reference.
The mean target concentration was determined to be 288.56 ng/mL in Balb/c Heparin plasma.
The mean target concentration was determined to be 327.73 ng/mL in C57BL/6 serum.
Serum levels in healthy individual mice
Duplicate measurements were performed on serum samples obtained from ten individual healthy mice. Data are reported as dilution factor-corrected interpolated values. The resultant mean Mouse Complement C5a concentration was 106.7 ng/mL, with a distribution spanning 37.24 to 366.38 ng/mL across the mouse cohort.
Intra-Assay Precision (Precision within an assay)
Three samples of known concentration were tested fifteen times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays).
Three samples of known concentration were tested in separate assays to assess inter-assay precision. Assays were performed with at least three lots of components.
Determination of Minimum Detectable Dose (MDD)
The MDD was determined using three independent lots of assay components. For each lot, 19 replicate measurements of the diluent (zero calibrator) were performed. The mean (AVERAGE) and standard deviation (STDEV) of the 19 replicates were calculated. The MDD for each lot was then calculated according to the following formula:
MDD = 2 × STDEV + AVERAGE
Protocol Diagram
