Multiplex Assay Kit for Complement Component 4a (C4a) ,etc. by FLIA (Flow Luminescence Immunoassay)
C4a anaphylatoxin
(Note: Up to 8-plex in one testing reaction)
- Product No.LMA389Mu
- Organism SpeciesMus musculus (Mouse) Same name, Different species.
- Sample TypeSerum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
- Test MethodDouble-antibody Sandwich
- Assay Length3.5h
- Detection Range1.95-2000pg/mL
- SensitivityThe minimum detectable dose of this kit is typically less than 0.65 pg/mL.
- Downloadn/a
- UOM 8Plex 7Plex 6Plex 5Plex 4Plex 3Plex 2Plex1Plex
- FOB
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Result
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Specificity
This assay has high sensitivity and excellent specificity for detection of Complement Component 4a (C4a) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Complement Component 4a (C4a) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.
Recovery
Matrices listed below were spiked with certain level of recombinant Complement Component 4a (C4a) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Complement Component 4a (C4a) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.
Matrix | Recovery range (%) | Average(%) |
serum(n=5) | 89-103 | 101 |
EDTA plasma(n=5) | 80-88 | 85 |
heparin plasma(n=5) | 83-93 | 87 |
sodium citrate plasma(n=5) | 86-99 | 91 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Complement Component 4a (C4a) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Complement Component 4a (C4a) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%
Linearity
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Complement Component 4a (C4a) ,etc. by FLIA (Flow Luminescence Immunoassay) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample | 1:2 | 1:4 | 1:8 | 1:16 |
serum(n=5) | 90-99% | 85-103% | 88-102% | 78-105% |
EDTA plasma(n=5) | 78-91% | 97-104% | 84-104% | 88-101% |
heparin plasma(n=5) | 83-93% | 88-96% | 79-96% | 79-98% |
sodium citrate plasma(n=5) | 89-96% | 92-99% | 87-94% | 86-95% |
Stability
The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.
Reagents and materials provided
Reagents | Quantity | Reagents | Quantity |
96-well plate | 1 | Plate sealer for 96 wells | 4 |
Pre-Mixed Standard | 2 | Standard Diluent | 1×20mL |
Pre-Mixed Magnetic beads (22#:C4a) | 1 | Analysis buffer | 1×20mL |
Pre-Mixed Detection Reagent A | 1×120μL | Assay Diluent A | 1×12mL |
Detection Reagent B (PE-SA) | 1×120μL | Assay Diluent B | 1×12mL |
Sheath Fluid | 1×10mL | Wash Buffer (30 × concentrate) | 1×20mL |
Instruction manual | 1 |
Assay procedure summary
1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.
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Magazine | Citations |
Journal of Vascular Surgery | Circulation levels of acute phase proteins in patients with Takayasu arteritis PubMed: 20100644 |
PLoS ONE | Identification of Altered Plasma Proteins by Proteomic Study in Valvular Heart Diseases and the Potential Clinical Significance PubMed: PMC3754973 |
PLoS One | Higher Serum Angiotensinogen Is an Indicator of IgA Vasculitis with Nephritis Revealed by Comparative Proteomes Analysis PubMed: 26098644 |
PLoS One | Plasma Protein Biomarkers of Hepatocellular Carcinoma in HCV-Infected Alcoholic Patients with Cirrhosis PubMed: 25789864 |
Experimental Eye Research | Quantitative analysis of hydroxyapatite-binding plasma proteins in genotyped individuals with late-stage age-related macular degeneration Pubmed:29580721 |
Diagnostics | Systemic Alterations of Immune Response-Related Proteins during Glaucoma Development in the Murine Model DBA/2J Pubmed: 32585848 |
Scientific Reports | Maternal Serum Proteomic Profiles of Pregnant Women With Type 1 Diabetes 34940989 |
Catalog No. | Related products for research use of Mus musculus (Mouse) Organism species | Applications (RESEARCH USE ONLY!) |
APA389Mu02 | Active Complement Component 4a (C4a) | Cell culture; Activity Assays. |
RPA389Mu01 | Recombinant Complement Component 4a (C4a) | Positive Control; Immunogen; SDS-PAGE; WB. |
RPA389Mu02 | Recombinant Complement Component 4a (C4a) | Positive Control; Immunogen; SDS-PAGE; WB. |
PAA389Mu02 | Polyclonal Antibody to Complement Component 4a (C4a) | WB; IHC; ICC; IP. |
PAA389Mu01 | Polyclonal Antibody to Complement Component 4a (C4a) | WB; IHC |
SEA389Mu | ELISA Kit for Complement Component 4a (C4a) | Enzyme-linked immunosorbent assay for Antigen Detection. |
LMA389Mu | Multiplex Assay Kit for Complement Component 4a (C4a) ,etc. by FLIA (Flow Luminescence Immunoassay) | FLIA Kit for Antigen Detection. |