Mini Samples ELISA Kit for Heat Shock Protein 70 (HSP70)

DnaK; 70 Kilodalton Heat Shock Proteins

Specificity

This assay has high sensitivity and excellent specificity for detection of Mini Samples Heat Shock Protein 70 (HSP70).
No significant cross-reactivity or interference between Mini Samples Heat Shock Protein 70 (HSP70) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Mini Samples Heat Shock Protein 70 (HSP70) and the recovery rates were calculated by comparing the measured value to the expected amount of Mini Samples Heat Shock Protein 70 (HSP70) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 98-105 102
EDTA plasma(n=5) 88-102 94
heparin plasma(n=5) 94-105 99

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Mini Samples Heat Shock Protein 70 (HSP70) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Mini Samples Heat Shock Protein 70 (HSP70) 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 Mini Samples Heat Shock Protein 70 (HSP70) 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) 78-102% 97-105% 83-91% 92-102%
EDTA plasma(n=5) 97-104% 84-98% 89-103% 93-103%
heparin plasma(n=5) 96-103% 95-102% 87-98% 90-101%

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
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 1×60µL Assay Diluent A 1×6mL
Detection Reagent B 1×60µL Assay Diluent B 1×6mL
TMB Substrate 1×4.5mL Stop Solution 1×3mL
Wash Buffer (30 × concentrate) 1×10mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 25µL standard or sample to each well. Incubate 1 hour at 37°C;
3. Aspirate and add 25µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 25µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 25µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 20µL Stop Solution. Read at 450nm immediately.

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Magazine Citations
Journal of Molecular Medicine Proteomic analysis reveals significant elevation of heat shock protein 70 in patients with chronic heart failure due to arrhythmogenic right ventricular cardiomyopathy SpringerLink: b8738j3571287425
Journal of Neuroinflammation Neuroinflammation in autism spectrum disorders PubMed: PMC3549857
Eur Arch Psychiatry Clin Neurosci. Luteal serum BDNF and HSP70 levels in women with premenstrual dysphoric disorder Pubmed: 23455589
Arch Bronconeumol Concentraciones de proteínas de estrés térmico, L-arginina y dimetilarginina asimétrica en pacientes con síndrome de apnea obstructiva del sue PubMed: 26055465
Arch Bronconeumol Heat Shock Proteins, PubMed: 26055465
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
Transplant Proc L-Alanyl-Glutamine Attenuates Oxidative Stress in Liver Transplantation Patients PubMed: 26518955
Plos One Long-Term Effects of Induced Hypothermia on Local and Systemic Inflammation-Results from a Porcine Long-Term Trauma Model Pubmed:27144532
J Pak Med Assoc. Elevated interictal serum HSP-70 levels as an indicator of neurodegeneration for chronic migraine mnstemps:33-1429777310
Egyptian Poultry Science Journal STUDY THE RELATIONSHIP BETWEEN SELENIUM AND HEAT SHOCK PROTEINS UNDER HEAT STRESS FOR LOCAL SINAI CHICKENS STRAIN abstract:3d115259031
Physiological consequences for rainbow trout (Oncorhynchus mykiss) of short-term exposure to increased suspended solid load S0144860916301935
Animal Science Journal Prokaryotic expression of chicken interferon-γ fusion protein and its effect on expression ofpoultry heat shock protein 70 under heat stress. pubmed:27753175
Pharmacol Rep High-dose testosterone enanthate supplementation boosts oxidative stress, but exerts little effect on the antioxidant barrier in sedentary adolescent male rat liver. pubmed:28531762
Biomed Pharmacother. SIRT1 inhibits releases of HMGB1 and HSP70 from human umbilical vein endothelial cells caused by IL-6 and the serum from a preeclampsia patient and protects the cells from death. pubmed:28122311
Immunobiology Concentrations of SP-A and HSP70 are associated with polarization of macrophages in pleural effusions of non-small cell lung cancer pubmed:29111316
Egyptian Poultry Science Journal EFFECT OF DIETARY SOURCE OF SELENIUM ON PRODUCTIVE AND REPRODUCTIVE PERFORMANCE OF SINAI LAYING HENS UNDER HEAT STRESS CONDITIONS. 125788660
Biomedicine & Pharmacotherapy SIRT1 inhibits releases of HMGB1 and HSP70 from human umbilical vein endothelial cells caused by IL-6 and the serum from a preeclampsia patient and protects the cells from death DOI:10.1016/j.biopha.2017.01.087
Egyptian Poultry Science Journal IMPACT OF CHROMIUM PICOLINATE SUPPLEMENTATION ON PRODUCTIVE PERFORMANCE, IMMUNE RESPONSE AND HEAT SHOCK PROTEINS OF BROILER CHICKENS UNDER HEAT-STRESS CONDITION. ISSN: 1110-5623
Aquaculture Physiological consequences of chronic exposure of rainbow trout (Oncorhynchus mykiss) to suspended solid load in recirculating aquaculture systems 10.1016/j.aquaculture.2017.11.030
Journal of dairy research Detection of 70 kDa heat shock protein in the saliva of dairy cows. pubmed:28625184
Journal of animal science Reducing rumen starch fermentation of wheat with three percent sodium hydroxide has the potential to ameliorate the effect of heat stress in grain-fed wethers pubmed:29293755
Biochemistry and Cell Biology Ectopic expression of Miro 1 ameliorates seizures and inhibits hippocampal neurodegeneration in a mouse model of pilocarpine epilepsy Pubmed:29365285
Egyptian Poultry Science Journal THE EFFECT OF CHROMIUM PICOLINATE SUPPLEMENTATION ON IMMUNE RESPONSE AND HEAT SHOCK PROTEINS OF LAYING HENS UNDER HEAT … article_5570_5c06907ac09a7724a58d2b796fe91aba.pdf
Egyptian Poultry Science Journal EFFECT OF MITIGATION OF HEAT STRESS BY EARLY HEATE ACCLIMATION AND GLUTAMINE INJECTION ON SOME PHYSIOLOGICAL MEASUREMENTS … :
FASEB Journal Signature-oriented investigation of the efficacy of multicomponent drugs against heart failure Pubmed: 30230922
Journal of Biological Sciences Maged Mohamed Abd-Elhakim Barakat, Noha Ahmed El-Boghdady, Ebtehal Kamal Eldin Farrag, Ataa Abdelhalim Said and Sylvia Edward Shaker
Biological Procedures Online Effective Visualization and Easy Tracking of Extracellular Vesicles in Glioma Cells
Journal of Biological Sciences Protective and Curative Effects of Bombax ceiba Flower and Ziziphus spina christi Fruit Extracts on Gastric Ulcer
博士论文 PHYSIOLOGICAL ADAPTATIONS TO HEAT ACCLIMATION; REPERCUSSIONS ON CYCLING PERFORMANCE
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