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ELISA Kit for Heat Shock Protein 27 (Hsp27)

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;

Specificity

This assay has high sensitivity and excellent specificity for detection of Heat Shock Protein 27 (Hsp27).
No significant cross-reactivity or interference between Heat Shock Protein 27 (Hsp27) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Heat Shock Protein 27 (Hsp27) and the recovery rates were calculated by comparing the measured value to the expected amount of Heat Shock Protein 27 (Hsp27) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 84-92 88
EDTA plasma(n=5) 84-102 88
heparin plasma(n=5) 90-97 94

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Heat Shock Protein 27 (Hsp27) 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) 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) 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) 83-101% 81-98% 88-104% 78-93%
EDTA plasma(n=5) 80-95% 95-103% 89-104% 86-101%
heparin plasma(n=5) 78-93% 90-104% 81-94% 86-98%

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
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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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
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