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Multiplex Assay Kit for Heat Shock Protein 27 (Hsp27) ,etc. by FLIA (Flow Luminescence Immunoassay)
HSPB1; HSP-B1; CMT2F; HSP28; Hsp25; Heat Shock 27kDa Protein 1; 28 kDa heat shock protein; Heat shock protein beta-1; Estrogen-regulated 24 kDa protein;
(Note: Up to 8-plex in one testing reaction)
- Product No.LMA693Hu
- Organism SpeciesHomo sapiens (Human) Same name, Different species.
- Sample TypeSerum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
- Test MethodDouble-antibody Sandwich
- Assay Length3.5h
- Detection Range0.05-50ng/mL
- SensitivityThe minimum detectable dose of this kit is typically less than 0.017 ng/mL.
- DownloadInstruction Manual
- UOM 8Plex 7Plex 6Plex 5Plex 4Plex 3Plex 2Plex1Plex
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Result
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Specificity
This assay has high sensitivity and excellent specificity for detection of Heat Shock Protein 27 (Hsp27) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Heat Shock Protein 27 (Hsp27) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.
Recovery
Matrices listed below were spiked with certain level of recombinant Heat Shock Protein 27 (Hsp27) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Heat Shock Protein 27 (Hsp27) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 91-103 | 98 |
| EDTA plasma(n=5) | 99-105 | 102 |
| heparin plasma(n=5) | 85-102 | 98 |
| sodium citrate plasma(n=5) | 97-105 | 102 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Heat Shock Protein 27 (Hsp27) ,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 Heat Shock Protein 27 (Hsp27) ,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 Heat Shock Protein 27 (Hsp27) ,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) | 80-94% | 80-101% | 95-105% | 78-90% |
| EDTA plasma(n=5) | 80-103% | 95-104% | 80-103% | 91-98% |
| heparin plasma(n=5) | 95-104% | 84-99% | 84-101% | 89-103% |
| sodium citrate plasma(n=5) | 96-105% | 94-105% | 85-94% | 80-92% |
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#:Hsp27) | 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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| Magazine | Citations |
| Animal Science Journal | The effect of heat stress on gene expression and synthesis of heat‐shock and milk proteins in bovine mammary epithelial cells PubMed: 26467738 |
| Nature Communications | HSP27 is a partner of JAK2-STAT5 and a potential therapeutic target in myelofibrosis Pubmed:29650953 |
| Advances in Experimental Medicine and Biology | Molecules of Damage-Associated Patterns in Bronchoalveolar Lavage Fluid and Serum in Chronic Obstructive Pulmonary Disease Pubmed:29429028 |
| Diabetes Research and Clinical Practice | The beneficial effects of 15 units of high-intensity circuit training in women is modified by age, baseline insulin resistance and physical capacity Pubmed: 31102684 |
| Journal of Immunology Research | Damage-Associated Molecular Patterns and Myeloid-Derived Suppressor Cells in Bronchoalveolar Lavage Fluid in Chronic Obstructive Pulmonary Disease Patients |
| Journal of Clinical Medicine | Damage-Associated Molecular Patterns and Th-Cell-Related Cytokines Released after Progressive Effort Pubmed: 32210109 |
| Medicine | Regulatory T cells, damage-associated molecular patterns, and myeloid-derived suppressor cells in bronchoalveolar lavage fluid interlinked with chronic obstructive … Pubmed:35687771 |
| Beilstein Archives | Immunization of mice with gold nanoparticles conjugated to thermostable tumor antigens prevents tumor development during transplantation |