ELISA Kit for Heme Oxygenase 1, Decycling (HO1)

HMOX1; Hsp32; HMOX1D

Specificity

This assay has high sensitivity and excellent specificity for detection of Heme Oxygenase 1, Decycling (HO1).
No significant cross-reactivity or interference between Heme Oxygenase 1, Decycling (HO1) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Heme Oxygenase 1, Decycling (HO1) and the recovery rates were calculated by comparing the measured value to the expected amount of Heme Oxygenase 1, Decycling (HO1) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 89-103 97
EDTA plasma(n=5) 87-101 97
heparin plasma(n=5) 85-101 89

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Heme Oxygenase 1, Decycling (HO1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Heme Oxygenase 1, Decycling (HO1) 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 Heme Oxygenase 1, Decycling (HO1) 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% 90-98% 84-92% 88-103%
EDTA plasma(n=5) 78-98% 89-99% 93-101% 98-105%
heparin plasma(n=5) 81-94% 91-103% 94-101% 90-98%

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
The American Journal of Pathology Characterization of the Intrarenal Renin-Angiotensin System in Experimental Alport Syndrome PubMed: 25777063
BioMed Research International Serum Heme Oxygenase-1 and BMP-7 Are Potential Biomarkers for Bone Metabolism in Patients with Rheumatoid Arthritis and Ankylosing Spondylitis Pubmed:27314037
Cancer Medicine Circulating CD14+HLA-DR-/low myeloid-derived suppressor cells in leukemia patients with allogeneic hematopoietic stem cell transplantation: novel clinical potential strategies for the prevention and cellular therapy of graft-versus-host disease Pubmed:27109254
Redox Report: Communications in Free Radical Research Decreased expression of heme oxygenase is associated with depressive symptoms and may contribute to depressive and hypertensive comorbidity Pubmed:26824276
Canadian Journal of Physiology and Pharmacology Sulforaphane protects against sodium valproate–induced acute liver injury pubmed:28112972
Peptides. Carnosine ameliorates cognitive deficits in streptozotocin-induced diabetic rats: Possible involved mechanisms. pubmed:27777064
european journal of pharmacology S-allyl cysteine ameliorates cognitive deficits in streptozotocin-diabetic rats via suppression of oxidative stress, inflammation, and acetylcholinesterase. pubmed:27887948
brain research Ellagic acid exerts protective effect in intrastriatal 6-hydroxydopamine rat model of Parkinson's disease: Possible involvement of ERβ/Nrf2/HO-1 signaling. pubmed:28238669
biomedicine & pharmacotherapy Citrus leaf extract reduces blood pressure and vascular damage in repeatedly heated palm oil diet-Induced hypertensive rats. pubmed:28068636
Inflammopharmacology. Implication of Nrf2/HO-1 pathway in the coloprotective effect of coenzyme Q10 against experimentally induced ulcerative colitis. pubmed:28050757
Environmental Toxicology and Pharmacology Ameliorative potential of linagliptin and/or calcipotriol on bleomycin-induced lung fibrosis: In vivo and in vitro study pubmed:28192751
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