Multiplex Assay Kit for Inhibitory Subunit Of NF Kappa B Alpha (IkBa) ,etc. by FLIA (Flow Luminescence Immunoassay)
NFKBIA; IKB-A; MAD-3; NFKBI; Nuclear Factor Of Kappa Light Polypeptide Gene Enhancer In B-Cells Inhibitor,Alpha; Major histocompatibility complex enhancer-binding protein MAD3
(Note: Up to 8-plex in one testing reaction)
- UOM
- FOB US$ 427.00 US$ 443.00 US$ 468.00 US$ 501.00 US$ 534.00 US$ 583.00 US$ 657.00 US$ 821.00
- Quantity
Overview
Properties
- Product No.LMB848Mu
- Organism SpeciesMus musculus (Mouse) Same name, Different species.
- ApplicationsFLIA Kit for Antigen Detection.
Research use only - DownloadInstruction Manual
- CategorySignal transductionTumor immunityHematologyDevelopmental science
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Recovery
Matrices listed below were spiked with certain level of recombinant Inhibitory Subunit Of NF Kappa B Alpha (IkBa) and the recovery rates were calculated by comparing the measured value to the expected amount of Inhibitory Subunit Of NF Kappa B Alpha (IkBa) in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 98-105 | 102 |
| EDTA plasma(n=5) | 89-104 | 93 |
| heparin plasma(n=5) | 96-103 | 101 |
| sodium citrate plasma(n=5) | 95-104 | 101 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Inhibitory Subunit Of NF Kappa B Alpha (IkBa) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Inhibitory Subunit Of NF Kappa B Alpha (IkBa) 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 Inhibitory Subunit Of NF Kappa B Alpha (IkBa) 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) | 88-101% | 88-101% | 78-89% | 78-93% |
| EDTA plasma(n=5) | 87-95% | 90-98% | 97-104% | 93-102% |
| heparin plasma(n=5) | 91-105% | 82-89% | 81-105% | 91-104% |
| sodium citrate plasma(n=5) | 85-96% | 81-96% | 90-98% | 81-101% |
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#:IkBa) | 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. Add 200μL analysis buffer solution to each well of the plate for pre-wetting;
2. Preparation of standards, reagents and samples before the experiment;
3. Add 100μL standard or sample to each well, add 10μL magnetic beads, and incubate 120 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60 minutes protect from light at 25°C on shaker;
5. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
6. Wash plate once on magnetic frame;
7. Add 100μL sheath fluid, swirl for 10 minutes, read on the machine.
Test principle
Anti-Inhibitory Subunit Of NF Kappa B Alpha (IkBa) antibody was coated on the surface of magnetic beads, to make solid phase carrier. Standard or sample were added to the magnetic beads, respectively, in which Inhibitory Subunit Of NF Kappa B Alpha (IkBa) was bound to the antibody connected to the solid phase carrier, and then biotinylated anti-Inhibitory Subunit Of NF Kappa B Alpha (IkBa) antibodies were added. PE-labeled avidin was added. There is a positive correlation between the concentration of Inhibitory Subunit Of NF Kappa B Alpha (IkBa) in the sample and corresponding value of MFI (Median Fluorescence Intensity).
