Multiplex Assay Kit for Histone H4 (H4) ,etc. by CBA (Cytometric Bead Array)

Specificity

This assay has high sensitivity for detection of Histone H4 (H4) ,etc. by CBA (Cytometric Bead Array).
100% cross-reactivity of Histone H4 (H4) ,etc. by CBA (Cytometric Bead Array) was observed among .

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.

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Magazine Citations
PLoS One. N-Acetyl-Heparin Attenuates Acute Lung Injury Caused by Acid Aspiration Mainly by Antagonizing Histones in Mice Pubmed:24816808
Crit Care. Impact of plasma histones in human sepsis and their contribution to cellular injury and inflammation Pubmed:25260379
Biomed Res Int A Novel Anti-Histone H2 Monoclonal Antibody, SSV Monoclonal Antibody, Improves Lung Injury and Survival in a Mouse Model of Lipopolysaccharide-Induced Sepsis-Like Syndrome PubMed: 25649891
Journal of Viral Hepatitis Patients with HBV-related acute-on-chronic liver failure have increased concentrations of extracellular histones aggravating cellular damage and systemic inflammation pubmed:27660136
Cell Death & Disease Dexmedetomidine inhibits astrocyte pyroptosis and subsequently protects the brain in in vitro and in vivo models of sepsis Pubmed: 30778043
Respir Res Histone H4 induces heparan sulfate degradation by activating heparanase in chlorine gas-induced acute respiratory distress syndrome Pubmed:35073921
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