Join Our Mailing List Today!

NOTICE: Online ordering will be disabled 1/27-2/3. Learn More.

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.

© 2024 FUJIFILM Irvine Scientific. All rights reserved.