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.