High Sensitive ELISA Kit for Inhibin Beta E (INHbE)

INH-bE; Activin beta-E chain

Specificity

This assay has high sensitivity and excellent specificity for detection of High Sensitive Inhibin Beta E (INHbE).
No significant cross-reactivity or interference between High Sensitive Inhibin Beta E (INHbE) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant High Sensitive Inhibin Beta E (INHbE) and the recovery rates were calculated by comparing the measured value to the expected amount of High Sensitive Inhibin Beta E (INHbE) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 86-103 98
EDTA plasma(n=5) 78-95 89
heparin plasma(n=5) 89-96 92

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level High Sensitive Inhibin Beta E (INHbE) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level High Sensitive Inhibin Beta E (INHbE) 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 High Sensitive Inhibin Beta E (INHbE) 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) 89-96% 94-101% 93-103% 78-98%
EDTA plasma(n=5) 82-92% 92-101% 80-101% 78-102%
heparin plasma(n=5) 93-101% 87-101% 82-104% 98-105%

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
Mitochondrion GDF15 is a novel biomarker to evaluate efficacy of pyruvate therapy for mitochondrial diseases Pubmed:25446397
PLoS One Inhibin βE (INHBE) is a possible insulin resistance-associated hepatokine identified by comprehensive gene expression analysis in human liver biopsy … Pubmed:29596463
Cell Reports Activin E Controls Energy Homeostasis in Both Brown and White Adipose Tissues as a Hepatokine Pubmed: 30380411
International Journal of Pharmacy and Biological Sciences Anti-Obesity Efficacy of Selected Plants Against Progesterone Induced Obesity
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