Mature BDNF ELISA Kit Wako, High Sensitive
- for Immunochemistry
- Manufacturer :
- FUJIFILM Wako Pure Chemical Corporation
- Storage Condition :
- Keep at 2-10 degrees C.
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96Tests
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Kit component
Kit component
| Antibody-coated Plate | 1 plate |
|---|---|
| Mature BDNF Standard (Lyophilized) | 1 vial |
| Buffer | 60 mL |
| Biotin-conjugated Antibody (Lyophilized) | 1 vial |
| Peroxidase-conjugated Streptavidin Solution | 100 μL |
| Luminescent Reagent 1 | 6 mL |
| Luminescent Reagent 2 | 6 mL |
| Wash Solution (10 x) | 100 mL |
| Plate Seal | 4 sheets |
Product Overview
Brain-derived neurotrophic factor (BDNF), which belongs to the nerve growth factor (NGF) family, is involved in neurogenesis, synaptogenesis, long-term potentiation etc. and is considered as a candidate marker for psychiatric diseases, including depression.
proBDNF, the precursor of BDNF, is processed to yield mature BDNF (mBDNF). Since proBDNF and mBDNF show completely different effects, there is a need for a method that can accurately measure mBDNF.
Mature BDNF ELISA Kit Wako, High Sensitive is a sandwich ELISA that combines anti mBDNF N-terminal end-specific monoclonal antibody and anti-BDNF monoclonal antibody. The cross-reactivity with proBDNF is substantially reduced by using end-specific antibodies, and the sensitivity is increased through use of a luminescent substrate. This kit can be used for detection mBDNF in human saliva and mouse serum or plasma.
Notice
*290-85801 Mature BDNF ELISA Kit Wako, High Sensitive, is an upgraded kit and replaces 298-83901 Mature BDNF Kit Wako, High Sensitive.
Features
- Specifically detects mBDNF
- Reactivity with human proBDNF: 1.30% - Highly sensitive
- lower limit of calibration curve: 0.116 pg/mL
Table 1: Sensitivity and cross-reactivity of BDNF ELISA kits
| Fujifilm Wako (Luminescence) |
Fujifilm Wako (Chromogenic) |
Company A | Company B | Company C | |
|---|---|---|---|---|---|
| Sensitivity lower limit of calibration curve |
0.116 pg/mL | 4.1 pg/mL | 62.5 pg/mL | 15.6 pg/mL | 15.0 pg/mL |
| Cross-Reactivity with human proBDNF |
1.30 % | Approx. 10 % | Approx. 10 % | Approx. 15 % | Approx. 50 % |
Principle
This kit is a sandwich ELISA that combines anti mBDNF N-terminal end-specific monoclonal antibody and anti-BDNF monoclonal antibody. The cross-reactivity with proBDNF is substantially reduced by using end-specific antibodies, and the sensitivity is increased through use of a luminescent substrate.
Kit Performance
| Calibration curve range | 0.116 - 50 pg/mL |
|---|---|
| Assay target | Mature BDNF |
| Analysis sample | Mouse serum/plasma/brain lysate Rat serum/plasma Human serum/plasma/saliva |
| Sample volume | 13 μL (4-fold dilution) |
| Measurement duration | Approx. 4 hours |
| Detection method | Luminescence (A plate reader for luminescence measurement is required) |
Example of Calibration Curve
Data
Spiked recovery test
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Human
Amount spiked(pg/mL) Measured value(pg/mL) Recovery volume(pg/mL) Recovery rate (%) Saliva 0.00 0.386 － － 0.500 0.875 0.489 97.8 5.00 5.47 5.08 102 30.0 30.1 29.7 99.0 Average 99.6 Serum 0.00 5.74 - - 0.500 6.21 0.470 94.0 5.00 10.9 5.16 103 30.0 35.7 30.0 100 Average 99.0 Plasma 0.00 4.27 - - 0.500 4.78 0.510 102 5.00 9.53 5.26 105 30.0 34.4 30.1 100 Average 102 -
Mouse
Amount spiked(pg/mL) Measured value(pg/mL) Recovery volume(pg/mL) Recovery rate (%) Serum 0.00 0.646 － － 0.500 1.11 0.464 92.8 5.00 5.38 4.73 94.6 30.0 30.7 30.1 100 Average 95.8 Plasma 0.00 0.116 － － 0.500 0.614 0.498 99.6 5.00 4.78 4.66 93.2 30.0 32.2 32.1 107 Average 99.9 Rat
Amount spiked(pg/mL) Measured value(pg/mL) Recovery volume(pg/mL) Recovery rate (%) Serum 0.00 0.860 － － 0.500 1.33 0.470 94.0 5.00 6.03 5.17 103 30.0 28.5 27.6 92.0 Average 96.3 Plasma 0.00 0.767 － － 0.500 1.26 0.493 98.6 5.00 5.95 5.18 104 30.0 28.4 27.6 92.0 Average 98.2
Dilution linearity test
Human
Mouse
Rat
Reactivity with similar proteins
| Species | Proteins | Reactivity (%) |
|---|---|---|
| Human | proBDNF | 1.30 |
| NGFβ | < 0.232 | |
| NT-3 | < 0.232 | |
| NT-4 | < 0.232 | |
| Mouse | proBDNF | 0.328 |
| NGFβ | < 0.232 | |
| NT-3 | < 0.232 | |
| NT-4 | < 0.232 |
Reactivity with proBDNF and the neurotrophin family (NGF, NT-3, and NT-4) was determined.
[Result]
Very low reactivity with proBDNF and the neurotrophin family (NGF, NT-3, NT-4) was confirmed
BDNF Measurement in brain lysate in wild-type and BDNF knockout mice
To confirm the specificity for BDNF, the kit was used to measure mBDNF in brain lysate collected from wild-type and BDNF knockout mice.
[Result]
The mBDNF level was very low in BDNF knockout mice, demonstrating the specificity for BDNF.
References
- Want, A., Morgan, J. E. and Barde, Y. A.: Sci. Rep., 13(1), 7740(2023).
Brain-derived neurotrophic factor measurements in mouse serum and plasma using a sensitive and specific enzyme-linked immunosorbent assay - Want, A. et al.: Brain Commun., 5(2), fcad046 (2023).
Brain-derived neurotrophic factor released from blood platelets prevents dendritic atrophy of lesioned adult central nervous system neurons - Ikenouchi, A. et al. : Brain Behav.,e3278(2023).
Association between salivarymature brain-derived neurotrophic factor and psychological distress in healthcare workers
Note: 1) - 3) use the predecessor of this kit, Mature BDNF ELISA Kit Wako, High Sensitive (Code No. 298-83901, discontinued) is used.
FAQ
About sample
- How should samples be stored?
- For long-term storage of samples, freezing at -80°C or below is recommended.
- What is the recommended method of collection of saliva samples?
- The passive drool method is recommended for the collection of saliva samples. The passive drool method requires subjects to salivate into a vial or tube.
It has been confirmed that samples collected using cotton swabs or inert polymer swabs can be used similarly to samples collected by the passive drool method. However, caution should be taken, because some swab materials may absorb target analytes.
- What should I do if I could not get a good result with a saliva sample?
- Thick saliva may not give expected results. In such a case, remove insoluble substances, such as mucin, from the saliva sample by freezing and thawing once, followed by centrifugation (example: 1,500 x g for 15 min), and use the supernatant for the assay.
- Which extraction buffer is recommended for the analysis of mouse brain lysate?
- Use RIPA buffer containing a protease inhibitor. To eliminate the influence of surfactant components in the buffer, dilute it with the buffer included in the kit, by a factor of at least twice.
- What is the recommended dilution ratio for each sample?
- The recommended dilution ratio is as follows. It is recommended that you optimize the dilution ratio to fall within the range of standard curve concentrations.
Sample Dilution ratio Mouse serum and plasma 4-fold Mouse brain lysate 2-320-fold Rat serum 50-fold Rat plasma 30-fold Human serum 10-2,000-fold Human plasma 10-80-fold Human saliva 2-8-fold
About the kit
- What reagents, instruments, and equipment are required for the assay using this kit?
- Purified water (distilled water)
- Test tubes for dilution of standard solution and samples
- Glass utensils for dilution of Wash Solution (e.g. volumetric cylinders and beakers)
- Pipettes with disposable tips (one capable of accurately pipetting 10μL of liquid, and one capable of accurately pipetting 200-500 μL of liquid)
- Continuous dispensing pipettes for 100 μL of liquid
- Water absorbents such as paper towels (used to remove the liquid remaining on the washed microplate)
- Stirrer (vortex type)
- Microplate shaker (approximately 600 to 800 rpm)
- Automatic washer (preferred) or washing bottles for 96-well microplates
- 96-well microplate reader for luminescence detection
- Software for data analysis
- Is it possible to purchase the antibodies provided in this kit?
- These antibodies are not sold separately.
- Can I divide plates?
- Since the plate in this kit are separable and each row (8 wells) is detachable, divided use of the kit is possible.
Overview / Applications
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Manufacturer Information
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For research use or further manufacturing use only. Not for use in diagnostic procedures.
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