ELISA Kit for Brain Derived Neurotrophic Factor (BDNF)

Neurotrophin; Abrineurin

Specificity

This assay has high sensitivity and excellent specificity for detection of Brain Derived Neurotrophic Factor (BDNF).
No significant cross-reactivity or interference between Brain Derived Neurotrophic Factor (BDNF) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Brain Derived Neurotrophic Factor (BDNF) and the recovery rates were calculated by comparing the measured value to the expected amount of Brain Derived Neurotrophic Factor (BDNF) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 78-101 86
EDTA plasma(n=5) 91-99 96
heparin plasma(n=5) 92-101 96

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Brain Derived Neurotrophic Factor (BDNF) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Brain Derived Neurotrophic Factor (BDNF) 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 Brain Derived Neurotrophic Factor (BDNF) 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) 91-98% 88-97% 96-103% 79-96%
EDTA plasma(n=5) 93-101% 79-104% 80-89% 78-104%
heparin plasma(n=5) 80-101% 92-99% 87-94% 90-97%

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
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The International Journal af Applied Research in Veterinary Medicine Effect of Sample Time in Clot Tube, Temperature, and Hemolysis Status on Canine Serum Brain-Derived Neurotrophic Factor (BDNF) Concentrations: Preliminary study articles:Vol14Iss3
J Chem Neuroanat. Both secreted and the cellular levels of BDNF attenuated due to tau hyperphosphorylation in primary cultures of cortical neurons. pubmed:27914953
Int Immunopharmacol. PPARr activation ameliorates postoperative cognitive decline probably through suppressing hippocampal neuroinflammation in aged mice. pubmed:27940378
The Journal of Maternal-Fetal & Neonatal Medicine Differential expression of cord blood neurotrophins in gestational diabetes: the impact of fetal growth abnormalities pubmed:28081639
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European Journal of Pharmacology Alpha-asarone attenuates depression-like behavior in nicotine-withdrawn mice: Evidence for the modulation of hippocampal pCREB levels during nicotine-withdrawal. pubmed:29042206
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REVISTA DA ASSOCIACAO MEDICA BRASILEIRA Physical Exercise Increased Brain-Derived Neurotrophic Factor in Elderly Population with Depression Pubmed: 30994834
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Transl Psychiatry Lithium augmentation of ketamine increases insulin signaling and antidepressant-like active stress coping in a rodent model of treatment-resistant depression 34824208
Iranian Journal of Basic Medical Sciences Adrenomedullin increases cAMP accumulation and BDNF expression in rat DRG and spinal motor neurons 34712429
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