Multiplex Assay Kit for Endothelial NOS (eNOS) ,etc. by FLIA (Flow Luminescence Immunoassay)

NOS3; ECNOS; NOS III; E-NOS; C-NOS; ENOS; CNOS; Constitutive NOS; Nitric Oxide Synthase 3, Endothelial

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

Specificity

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

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 80-98 93
EDTA plasma(n=5) 80-95 83
heparin plasma(n=5) 98-105 102

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Endothelial NOS (eNOS) ,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 Endothelial NOS (eNOS) ,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 Endothelial NOS (eNOS) ,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) 78-99% 97-104% 89-103% 84-92%
EDTA plasma(n=5) 90-103% 86-104% 78-93% 84-99%
heparin plasma(n=5) 99-105% 79-93% 85-104% 93-105%

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#:eNOS) 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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APA868Hu01 Active Endothelial NOS (eNOS) Cell culture; Activity Assays.
RPA868Hu03 Recombinant Endothelial NOS (eNOS) Positive Control; Immunogen; SDS-PAGE; WB.
RPA868Hu04 Recombinant Endothelial NOS (eNOS) Positive Control; Immunogen; SDS-PAGE; WB.
RPA868Hu01 Recombinant Endothelial NOS (eNOS) Positive Control; Immunogen; SDS-PAGE; WB.
PAA868Hu04 Polyclonal Antibody to Endothelial NOS (eNOS) WB; IHC; ICC; IP.
PAA868Hu03 Polyclonal Antibody to Endothelial NOS (eNOS) WB; IHC; ICC; IP.
PAA868Hu01 Polyclonal Antibody to Endothelial NOS (eNOS) WB; IHC; ICC; IP.
LAA868Hu71 Biotin-Linked Polyclonal Antibody to Endothelial NOS (eNOS) WB; IHC; ICC.
MAA868Hu22 Monoclonal Antibody to Endothelial NOS (eNOS) WB; IHC; ICC; IP.
SEA868Hu ELISA Kit for Endothelial NOS (eNOS) Enzyme-linked immunosorbent assay for Antigen Detection.
LMA868Hu Multiplex Assay Kit for Endothelial NOS (eNOS) ,etc. by FLIA (Flow Luminescence Immunoassay) FLIA Kit for Antigen Detection.