ELISA Kit for Heparin Binding Epidermal Growth Factor Like Growth Factor (HBEGF)

HB-EGF; DTR; DTS; DTSF; HEGFL; Diphtheria Toxin Receptor; Proheparin-binding EGF-like growth factor; Diphtheria toxin receptor

Specificity

This assay has high sensitivity and excellent specificity for detection of Heparin Binding Epidermal Growth Factor Like Growth Factor (HBEGF).
No significant cross-reactivity or interference between Heparin Binding Epidermal Growth Factor Like Growth Factor (HBEGF) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Heparin Binding Epidermal Growth Factor Like Growth Factor (HBEGF) and the recovery rates were calculated by comparing the measured value to the expected amount of Heparin Binding Epidermal Growth Factor Like Growth Factor (HBEGF) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 94-105 101
EDTA plasma(n=5) 96-103 99
heparin plasma(n=5) 78-97 89

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Heparin Binding Epidermal Growth Factor Like Growth Factor (HBEGF) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Heparin Binding Epidermal Growth Factor Like Growth Factor (HBEGF) 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 Heparin Binding Epidermal Growth Factor Like Growth Factor (HBEGF) 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) 91-105% 78-96% 93-101% 98-105%
EDTA plasma(n=5) 90-97% 86-104% 84-104% 82-96%
heparin plasma(n=5) 91-101% 81-105% 92-101% 84-102%

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
Neuropathology Astrocyte ERK phosphorylation precedes K+-induced swelling but follows hypotonicity-induced swelling. PubMed: 21118399
Toxicological Sciences Cypermethrin Induces Astrocyte Apoptosis by the Disruption of the Autocrine/Paracrine Mode of Epidermal Growth Factor Receptor Signaling Pubmed: source
PAIN RESEARCH 軽度炎症性膀胱痛マウスモデルの構築とNGFの関与 Jstage:Source
European Journal of Pain A novel method for assessing bladder-related pain reveals the involvement of nerve growth factor in painassociated with cyclophosphamide-induced chronic cystitis in mice. Pubmed:25820250
Kinki daigaku 2 型糖尿病モデルラットにおける糖尿病性腎症の進行とHB-EGF の関連性 17844
laboratory investigation Heparin-binding epidermal growth factor contributes to COPD disease severity by modulating airway fibrosis and pulmonary epithelial–mesenchymal transition Pubmed:29581578
International Journal of Biological Sciences CD9 regulates keratinocyte migration by negatively modulating the sheddase activity of ADAM17
Redox Biology Tumoral NOX4 recruits M2 tumor-associated macrophages via ROS/PI3K signaling-dependent various cytokine production to promote NSCLC growth Pubmed: 30769285
JOURNAL OF CELLULAR AND MOLECULAR MEDICINE Polarization of ADAM17‐driven EGFR signalling in electric field‐guided collective migration of epidermal sheets Pubmed: 33164313
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