Multiplex Assay Kit for Histone H4 (H4) ,etc. by CBA (Cytometric Bead Array) 
Overview
Properties
- Product No.CMA289Mi
- Organism Species Same name, Different species.
- ApplicationsFLIA Kit for Antigen Detection.
Research use only - DownloadInstruction Manual
- CategorySignal transductionDevelopmental science
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Packages (Simulation)
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Packages (Simulation)
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ISO9001: 2008, ISO13485: 2003 Registered
Recovery
Matrices listed below were spiked with certain level of recombinant Histone H4 (H4) ,etc. by CBA (Cytometric Bead Array) and the recovery rates were calculated by comparing the measured value to the expected amount of Histone H4 (H4) ,etc. by CBA (Cytometric Bead Array) in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 97-104 | 101 |
| EDTA plasma(n=5) | 78-95 | 84 |
| heparin plasma(n=5) | 90-97 | 94 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Histone H4 (H4) ,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 Histone H4 (H4) ,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 Histone H4 (H4) ,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) | 80-102% | 89-103% | 94-102% | 78-89% |
| EDTA plasma(n=5) | 97-104% | 85-103% | 78-101% | 79-91% |
| heparin plasma(n=5) | 82-99% | 85-104% | 86-101% | 92-101% |
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 |
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-Histone H4 (H4) ,etc. by CBA (Cytometric Bead Array) 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 Histone H4 (H4) ,etc. by CBA (Cytometric Bead Array) was bound to the antibody connected to the solid phase carrier, and then biotinylated anti-Histone H4 (H4) ,etc. by CBA (Cytometric Bead Array) antibodies were added. PE-labeled avidin was added. There is a positive correlation between the concentration of Histone H4 (H4) ,etc. by CBA (Cytometric Bead Array) in the sample and corresponding value of MFI (Median Fluorescence Intensity). Flow Cytometer was used to determine and calculate the sample concentration.
Giveaways
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Citations
- N-Acetyl-Heparin Attenuates Acute Lung Injury Caused by Acid Aspiration Mainly by Antagonizing Histones in MicePubmed:24816808
- Impact of plasma histones in human sepsis and their contribution to cellular injury and inflammationPubmed:25260379
- A Novel Anti-Histone H2 Monoclonal Antibody, SSV Monoclonal Antibody, Improves Lung Injury and Survival in a Mouse Model of Lipopolysaccharide-Induced Sepsis-Like SyndromePubMed: 25649891
- Patients with HBV-related acute-on-chronic liver failure have increased concentrations of extracellular histones aggravating cellular damage and systemic inflammationpubmed:27660136
- Dexmedetomidine inhibits astrocyte pyroptosis and subsequently protects the brain in in vitro and in vivo models of sepsisPubmed: 30778043
- Histone H4 induces heparan sulfate degradation by activating heparanase in chlorine gas-induced acute respiratory distress syndromePubmed:35073921