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Multiplex Assay Kit for Heparin Cofactor II (HCII) ,etc. by FLIA (Flow Luminescence Immunoassay)

SERPIND1; HC2; HCF2; HC II; HLS2; LS2; Serpin D1; Serpin Peptidase Inhibitor Clade D Member 1; Protease inhibitor leuserpin-2

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 80-101 82
EDTA plasma(n=5) 80-102 93
heparin plasma(n=5) 79-105 80
sodium citrate plasma(n=5) 80-97 87

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Heparin Cofactor II (HCII) ,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 Heparin Cofactor II (HCII) ,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 Heparin Cofactor II (HCII) ,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) 85-99% 89-101% 79-94% 83-96%
EDTA plasma(n=5) 88-102% 94-103% 90-102% 97-104%
heparin plasma(n=5) 90-98% 79-101% 92-99% 84-102%
sodium citrate plasma(n=5) 91-101% 80-95% 91-98% 82-102%

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#:HCII) 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 Immunology Proteolytic Activation Transforms Heparin Cofactor II into a Host Defense Molecule PubMed: PMC3677170
Biomacromolecules Anticoagulant properties of poly (sodium 2-(acrylamido)-2-methylpropanesulfonate)-based di-and triblock polymers Pubmed:29733637
Catalog No. Related products for research use of Homo sapiens (Human) Organism species Applications (RESEARCH USE ONLY!)
RPA284Hu01 Recombinant Heparin Cofactor II (HCII) Positive Control; Immunogen; SDS-PAGE; WB.
PAA284Hu01 Polyclonal Antibody to Heparin Cofactor II (HCII) WB; IHC; ICC; IP.
MAA284Hu22 Monoclonal Antibody to Heparin Cofactor II (HCII) WB; IHC; ICC; IP.
SEA284Hu ELISA Kit for Heparin Cofactor II (HCII) Enzyme-linked immunosorbent assay for Antigen Detection.
AEA284Hu ELISA Kit for Anti-Heparin Cofactor II Antibody (Anti-HCII) Enzyme-linked immunosorbent assay for Antibody Detection.
LMA284Hu Multiplex Assay Kit for Heparin Cofactor II (HCII) ,etc. by FLIA (Flow Luminescence Immunoassay) FLIA Kit for Antigen Detection.