Can Get Signal - toyobo.co.jp

1 - 18 - 05-07 Can Get Signal immunoreaction enhancer Solution (Code No. NKB-101, NKB-101T, NKB-201, NKB-301) toyobo CO., LTD. Life Science Department OSAKA JAPAN A3302K - 0 - [1] 1 [2] 2 [3] 2 [4] 3 [5] 7 [6] ELISA 10 [7] 14 [8] 16 [9] 17 - 1 - [1]

2 Can Get Signal immunoreaction enhancer Solution ELISA(enzyme-linked immunosorbent assay) 1. Can Get Signal S/N 2. Can Get Signal ELISA 3. Ready-to-Use Can Get Signal Ready-to-Use - 2 - [2] NKB-101 NKB-101 TNKB-201 NKB-301 Solution 1 for primary antibody 250ml 50ml 250ml - Solution 2 for secondary antibody 250ml 50ml - 250ml Solution 1 Solution 2 4 C [3] TBS(TBS-T) PBS(PBS-T) Solution 1 Solution 2 (NKB-201 Solution 1 NKB-301 Solution 2 ) ( )

3 Solution 2 Solution 1 ELISA Solution 1 ([6] ELISA ) BSA ELISA - 3 - [4] SDS PVDF( ) (Horseradish Peroxidase: HRP) (Alkaline Phosphatase: AP) IgG ( ) ( ) (1) ( FAS-1000 ) X ( ) (2) (* [7] ) (HRP AP ) ( ) PVDF (Amersham Hybond-P ) * TBS-T* PBS-T* HRP (Amersham ECL Plus HRP ) AP (Promega Western Blue AP ) (3) SDS-PAGE PVDF - 4 - (4) PVDF ( )

4 PVDF PVDF PVDF 1. PVDF 2. PVDF 1 3. PVDF 5 4. PVDF 10 (5) PVDF ( ) 1. PVDF 8 2. 3. 4 SDS-PAGE PVDF 4 PVDF (x4 ) (x4 ) PVDF SDS-PAGE : - 5 - 4. PVDF (cm2) x mA 1 8 PVDF 5. TBS-T PVDF 5 TBS-T PBS-T ( ) PBS-T AP AP TBS-T (6) 1.

5 1g 20ml TBS-T ( ) BSA( ) - 2. PVDF 1 3. TBS-T 4. TBS-T 15 x1 5 x2 (7) 1. Can Get Signal Solution 1 ([5] ) Solution 2 Solution 1 (8) 2. PVDF - 6 - 3. TBS-T 10 x3 (8) 1. Can Get Signal Solution 2 ([5] ) 2.

6 PVDF 3. TBS-T 10 x3 (9) (HRP ) HRP 1. ECL Plus (Amersham ) Solution A B 40:1 PVDF (cm2) x ml 2. PVDF ECL 5 3. PVDF ECL ( ) 4. ( FAS-1000 ) X (10) (AP ) AP 1. PVDF Western Blue(Promega ) 37 C 15 PVDF (cm2) x ml 3. PVDF Western Blue ( FAS-III ) - 7 - [5] PVDF (DNA RNA ) PVDF (1) ( FAS-1000 ) X ( ) (2) (* [7] ) (HRP AP ) (Difco ) PVDF (Amersham Hybond-P )

7 TBS-T* PBS-T* HRP (Amersham ECL Plus HRP ) (3) PVDF ( ) 1. PVDF 2. PVDF 1 3. PVDF 5 - 8 - (4) ( ) . TBS PVDF 2 l (5) 1. 1g 20ml TBS-T [4] 2. PVDF 1 3. TBS-T 4. TBS-T 15 x1 5 x2 (6) 1. Can Get Signal Solution 1 100 500 1,000 5,000 10,000 Solution 2 Solution 1 (7) 2.

8 PVDF 1 6 3. 4. TBS-T 10 x3 6 - 9 - (7) 1. Can Get Signal Solution 2 2. 3. TBS-T 10 x3 (8) (HRP ) HRP 1. ECL Plus (Amersham ) Solution A B 40:1 PVDF (cm2) x ml 2. PVDF ECL 5 3. PVDF ECL ( ) 4. ( FAS-1000 ) X - 10 - [6] ELISA ELISA(Enzyme-linked Immunosorbent Assay) ELISA (1) (2) (3) ( ) (3) 3 (1) ELISA 8 12 ( ) (2) (* [7] )

9 ( ) (HRP AP ) (Difco ) 96 ELISA 96 50mM Carbonate Buffer, pH * TBS PBS* TBS-T* PBS-T* HRP (BioFX TMB HRP ) 1N 1N - 11 - (3) 1. 50mM Carbonate Buffer, 100 96 20 (10 g/ml) 2. 37 C 4 C 16 37 C 4 C 16 3.

10 96 4. PBS 250 3 1:201:401:801:160 1:3201:200 1:400 1:800 1:1600 1:3200 1:6400 : ( ) - 12 - (4) 1. 3% PBS ( ) BSA( ) 2. 250 3. 37 C 4 C 16 4 C 4.