Multiplex Assay Kit for Neurofilament, Light Polypeptide (NEFL) ,etc. by CBA (Cytometric Bead Array)
CMT1F; CMT2E; NF-L; NF68; NFL; 68 kDa neurofilament protein; Neurofilament triplet L protein
Overview
Properties
- Product No.CME038Ra
- Organism SpeciesRattus norvegicus (Rat) Same name, Different species.
- ApplicationsFLIA Kit for Antigen Detection.
Research use only - DownloadInstruction Manual
- CategoryNeuro science
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Recovery
Matrices listed below were spiked with certain level of recombinant Neurofilament, Light Polypeptide (NEFL) ,etc. by CBA (Cytometric Bead Array) and the recovery rates were calculated by comparing the measured value to the expected amount of Neurofilament, Light Polypeptide (NEFL) ,etc. by CBA (Cytometric Bead Array) in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 80-92 | 87 |
| EDTA plasma(n=5) | 79-101 | 91 |
| heparin plasma(n=5) | 98-105 | 102 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Neurofilament, Light Polypeptide (NEFL) ,etc. by CBA (Cytometric Bead Array) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Neurofilament, Light Polypeptide (NEFL) ,etc. by CBA (Cytometric Bead Array) 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 Neurofilament, Light Polypeptide (NEFL) ,etc. by CBA (Cytometric Bead Array) 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) | 97-105% | 81-95% | 87-97% | 89-103% |
| EDTA plasma(n=5) | 97-105% | 79-99% | 93-103% | 89-97% |
| heparin plasma(n=5) | 94-103% | 80-97% | 87-96% | 89-96% |
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#:NEFL) | 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 50μL standard or sample to each well, add 50μL prepared Detection Reagent A, add 10μL magnetic beads, and incubate 90 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
5. Wash plate once on magnetic frame;
6. Add 100μL sheath fluid, swirl for 10 minutes, read on the machine.
Test principle
Anti-Neurofilament, Light Polypeptide (NEFL) ,etc. by CBA (Cytometric Bead Array) antibody was coated on the surface of magnetic beads. A competitive inhibition reaction is launched between biotin labeled Neurofilament, Light Polypeptide (NEFL) ,etc. by CBA (Cytometric Bead Array) and unlabeled Neurofilament, Light Polypeptide (NEFL) ,etc. by CBA (Cytometric Bead Array) (Standards or samples) with the pre-coated antibody specific to Neurofilament, Light Polypeptide (NEFL) ,etc. by CBA (Cytometric Bead Array). PE-labeled avidin was added. The amount of bound PE conjugate is reverse proportional to the concentration of Neurofilament, Light Polypeptide (NEFL) ,etc. by CBA (Cytometric Bead Array) in the sample, the intensity of MFI developed is reverse proportional to the concentration of Neurofilament, Light Polypeptide (NEFL) ,etc. by CBA (Cytometric Bead Array) in the sample.
Giveaways
Increment services
Citations
- Effect of carotid endarterectomy on brain damage markersPubmed:27126899
- Effect of carotid endarterectomy on brain damage markers.pubmed:27126899
- Cytokines and biological markers in autoimmune GFAP astrocytopathy: The potential role for pathogenesis and therapeutic implicationsPubmed: 31254929
- Neurofilament Light Chain Is a Biomarker of Neurodegeneration in Ataxia Telangiectasia33893614
- Phase 1b dose-escalation, safety, and pharmacokinetic study of IC14, a monoclonal antibody against CD14, for the treatment of amyotrophic lateral sclerosis34678870
- Fatty Acid-Binding Protein 7 (FABP-7), Glutamic Acid and Neurofilament Light Chain (NFL) as Potential Markers of Neurodegenerative Disorders in Psoriatic Patients …Pubmed:35566558
