Human Complement fragment 3a,C3a ELISA Kit
Product Details
| Description |
The product CSB-E08509h is a ready-to-use microwell, strip plate ELISA Kit for quantitatively analyzing the amount of the Complement fragment 3a (C3a) in human serum, plasma, and tissue homogenates. C3a is the cleavage product of complement C3 during the activation of the complement cascade. It is traditionally regarded as an anaphylatoxin that leads to smooth muscle contraction, histamine from mast cells, and vasodilation. Additionally, C3a mediates numerous other biological processes, including inflammatory and anti-inflammatory responses, through the interaction with its receptor C3aR. C3a-C3aR signaling enhances pro-metastatic cytokine secretion and extracellular matrix components expression of carcinoma-associated fibroblasts (CAFs) via the activation of PI3K-AKT signaling, thus facilitating the metastasis of breast cancer. C3a/C3aR axis-mediated negative regulation of proinflammatory responses and skewing macrophages towards an M2-like phenotype is conducive to the protective
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| Target Name |
Complement fragment 3a,C3a |
| Alternative Names |
N/A |
| Abbreviation |
C3a |
| Species |
Homo sapiens (Human) |
| Sample Types |
serum, plasma, tissue homogenates |
| Detection Range |
31.25 ng/mL-2000 ng/mL |
| Sensitivity |
7.81 ng/mL |
| Assay Time |
1-5h |
| Sample Volume |
50-100ul |
| Detection Wavelength |
450 nm |
| Research Area |
Immunology |
| Assay Principle |
quantitative |
| Measurement |
Sandwich |
| Precision |
| Intra-assay Precision (Precision within an assay): CV% |
| Three samples of known concentration were tested twenty times on one plate to assess. |
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| Inter-assay Precision (Precision between assays): CV% |
| Three samples of known concentration were tested in twenty assays to assess. |
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| Linearity |
| To assess the linearity of the assay, samples were spiked with high concentrations of human C3a in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay. |
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Sample |
Serum(n=4) |
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| 1:1 |
Average % |
92 |
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| Range % |
88-99 |
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| 1:2 |
Average % |
95 |
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| Range % |
90-105 |
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| 1:4 |
Average % |
92 |
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| Range % |
82-105 |
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| 1:5 |
Average % |
95 |
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| Range % |
90-101 |
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| Recovery |
| The recovery of human C3a spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section. |
| Sample Type |
Average % Recovery |
Range |
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| Serum (n=5) |
97 |
95-100 |
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| EDTA plasma (n=4) |
98 |
95-102 |
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| Typical Data |
| These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed. |
| ng/ml |
OD1 |
OD2 |
Average |
Corrected |
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| 2000 |
2.769 |
2.777 |
2.773 |
2.597 |
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| 1000 |
2.374 |
2.381 |
2.378 |
2.202 |
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| 500 |
1.801 |
1.790 |
1.796 |
1.620 |
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| 250 |
1.041 |
1.094 |
1.068 |
0.892 |
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| 125 |
0.632 |
0.652 |
0.642 |
0.466 |
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| 62.5 |
0.461 |
0.473 |
0.467 |
0.291 |
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| 31.25 |
0.259 |
0.284 |
0.272 |
0.096 |
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| 0 |
0.175 |
0.176 |
0.176 |
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| Materials provided |
- A micro ELISA plate --- The 96-well plate has been pre-coated with an anti-human C3a antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
- Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
- One vial Biotin-labeled C3a antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
- One vial HRP-avidin (100 x concentrate) (120 μl/bottle) ---Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
- One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the Biotin-antibody.
- One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
- One vial Sample Diluent (50 ml/bottle)---Dilute the sample to an appropriate concentration.
- One vial Wash Buffer (25 x concentrate) (20 ml/bottle) ---Wash away unbound or free substances.
- One vial TMB Substrate (10 ml/bottle) ---Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
- One vial Stop Solution (10 ml/bottle) ---Stop the color reaction. The solution color immediately turns from blue to yellow.
- Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
- An instruction manual
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| Materials not provided |
- A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
- An incubator can provide stable incubation conditions up to 37°C±5°C.
- Centrifuge
- Vortex
- Squirt bottle, manifold dispenser, or automated microplate washer
- Absorbent paper for blotting the microtiter plate
- 50-300ul multi-channel micropipette
- Pipette tips
- Single-channel micropipette with different ranges
- 100ml and 500ml graduated cylinders
- Deionized or distilled water
- Timer
- Test tubes for dilutio
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| Storage |
Store at 2-8°C. Please refer to protocol. |
| Lead Time |
3-5 working days |
