Multiplex Assay Kit for Activated Protein C (APC) ,etc. by FLIA (Flow Luminescence Immunoassay)

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
EDTA plasma(n=5) 88-95 92
heparin plasma(n=5) 88-104 99

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Activated Protein C (APC) ,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 Activated Protein C (APC) ,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 Activated Protein C (APC) ,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
EDTA plasma(n=5) 88-104% 96-105% 95-103% 78-92%
heparin plasma(n=5) 98-105% 88-102% 94-102% 86-94%

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#:APC) 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
The Journal of Immunology Activated Protein C Attenuates Systemic Lupus Erythematosus and Lupus Nephritis in MRL-Fas(lpr) Mice Jimmunol: 3413
Critical Care Disseminated intravascular coagulation or acute coagulopathy of trauma shock early after trauma? An observational study BioMed: cc10553
Molecular Endocrinology Intraovarian Thrombin and Activated Protein C Signaling System Regulates Steroidogenesis during the Periovulatory Period Endo: source
Scandinavian Journal of Trauma, Resuscitation and Emergency Medicine High levels of soluble VEGF receptor 1 early after trauma are associated with shock, sympathoadrenal activation, glycocalyx degradation and inflammation in severely injured patients: a prospective study Sjtrem:1757-7241
Crit Care. Impact of plasma histones in human sepsis and their contribution to cellular injury and inflammation Pubmed:25260379
Am J Physiol Heart Circ Physiol. Monocytes regulate systemic coagulation and inflammation in abdominal sepsis Pubmed:25502108
Neuroscience Protective effects of thrombomodulin on microvascular permeability after subarachnoid hemorrhage in mouse model PubMed: 25936678
Journal of Intensive Care Activated protein C does not increase in the early phase of trauma with disseminated intravascular coaCavia (Guinea pig )lation: comparison with acute coaCavia (Guinea pig )lopathy of trauma-shock Pubmed:26734467
Journal of Biological Chemistry Brain microvascular endothelial cells exhibit lower activation of the alternative complement pathway than glomerular microvascular endothelial cells. JBC:Source
J Thromb Haemost A multicenter prospective validation study on disseminated intravascular coagulation in trauma‐induced coagulopathy Pubmed: 32480432
Validation of the Relationship Between Coagulopathy and Localization of Hydroxyethyl Starch on the Vascular Endothelium in a Rat Hemodilution Model 34021192
Catalog No. Related products for research use of Mus musculus (Mouse) Organism species Applications (RESEARCH USE ONLY!)
RPA738Mu01 Recombinant Activated Protein C (APC) Positive Control; Immunogen; SDS-PAGE; WB.
PAA738Mu01 Polyclonal Antibody to Activated Protein C (APC) WB; IHC; ICC; IP.
SEA738Mu ELISA Kit for Activated Protein C (APC) Enzyme-linked immunosorbent assay for Antigen Detection.
LMA738Mu Multiplex Assay Kit for Activated Protein C (APC) ,etc. by FLIA (Flow Luminescence Immunoassay) FLIA Kit for Antigen Detection.