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Mature BDNF ELISA Kit Wako, High Sensitive Data
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
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
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.