Multiplex Assay Kit for Complement Component 5 (C5) ,etc. by FLIA (Flow Luminescence Immunoassay)

CPAMD4; C5a anaphylatoxin; C3 and PZP-like alpha-2-macroglobulin domain-containing protein 4

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Complement Component 5 (C5) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Complement Component 5 (C5) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Complement Component 5 (C5) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Complement Component 5 (C5) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 90-102 96
EDTA plasma(n=5) 84-91 88
heparin plasma(n=5) 96-103 99
sodium citrate plasma(n=5) 88-102 93

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Complement Component 5 (C5) ,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 5 (C5) ,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 5 (C5) ,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) 92-99% 90-99% 82-101% 84-101%
EDTA plasma(n=5) 95-103% 93-105% 88-101% 95-105%
heparin plasma(n=5) 80-93% 88-103% 98-105% 97-104%
sodium citrate plasma(n=5) 96-104% 88-99% 96-104% 82-96%

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#:C5) 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 Clinical Immunology Complement activation contributes to the injury and outcome of kidney in human anti-glomerular basement membrane disease. PubMed: 22941511
Journal of Neuroimmunology Acute and prolonged complement activation in the central nervous system during herpes simplex encephalitis Pubmed:27235358
Cancer Letters Complement C5a/C5aR pathway potentiates the pathogenesis Q5 of gastric cancer by down-regulating p21 expression pubmed:29031586
Proteomics Clinical Applications iTRAQ‐Based Proteomic Analysis Reveals Protein Profile in Plasma from Children with Autism Pubmed:29274201
American journal of physiology. Endocrinology and metabolism Impaired insulin signaling in the B10.D2-Hc H2 H2-T18/oSnJ mouse model of complement factor 5 (C5) deficiency Pubmed: 31084499
Proteomics Clinical Applications Comparative proteomics analysis of serum proteins in gestational diabetes during early and middle stages of pregnancy Pubmed: 31162828
Aging C5aR deficiency attenuates the breast cancer development via the p38/p21 axis Pubmed: 32669478
Catalog No. Related products for research use of Homo sapiens (Human) Organism species Applications (RESEARCH USE ONLY!)
RPB969Hu01 Recombinant Complement Component 5 (C5) Positive Control; Immunogen; SDS-PAGE; WB.
APB969Hu02 Active Complement Component 5 (C5) Cell culture; Activity Assays.
RPB969Hu02 Recombinant Complement Component 5 (C5) Positive Control; Immunogen; SDS-PAGE; WB.
PAB969Hu01 Polyclonal Antibody to Complement Component 5 (C5) WB; IHC; ICC; IP.
PAB969Hu02 Polyclonal Antibody to Complement Component 5 (C5) WB; IHC; ICC; IP.
LAB969Hu71 Biotin-Linked Polyclonal Antibody to Complement Component 5 (C5) WB; IHC; ICC.
MAB969Hu22 Monoclonal Antibody to Complement Component 5 (C5) WB; IHC; ICC; IP.
SEB969Hu ELISA Kit for Complement Component 5 (C5) Enzyme-linked immunosorbent assay for Antigen Detection.
LMB969Hu Multiplex Assay Kit for Complement Component 5 (C5) ,etc. by FLIA (Flow Luminescence Immunoassay) FLIA Kit for Antigen Detection.