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w a s h i n g O p e r a t i o n w i t h 4 .
c o l o r A d d C h r o m o g e n S o l u t i o n A 5 0 u l a n d C h r o m o g e n S o l u t i o n B t o e a c h w e l l , I n c u b a t e f o r 1 5 m i n a t 3 7 !
S t o p t h e r e a c t i o n A d d S t o p S o l u t i o n 5 0 l t o e a c h w e l l , S t o p t h e r e a c t i o n ( t h e b l u e c o l o r c h a n g e t o y e l l o w c o l o r I m m e d i a t e l y ) .
a s s a y t a k e b l a n k w e l l a s z e r o , m e a s u r e t h e o p t i c a l d e n s i t ( O D ) a t 4 5 0 n m a f t e r A d d i n g S t o p S o l u t i o n a n d w i t h i n 1 5 m i n .
C a l c u l a t e
T a k e t h e s t a n d a r d d e n s i t y a s t h e h o r i z o n t a l , t h e O D v a l u e f o r t h e v e r t i c a l , d r a w t h e s t a n d a r d c u r v e o n g r a p h p a p e r , F i n d o u t t h e c o r r e s p o n d i n g d e n s i t y a c c o r d i n g t o t h e s a m p l e O D v a l u e b y t h e S a m p l e c u r v e , m u l t i p l i e d b y t h e d i l u t i o n m u l t i p l e , o r c a l c u l a t e t h e s t r a i g h t l i n e r e g r e s s i o n e q u a t i o n o f t h e s t a n d a r d c u r v e w i t h t h e s t a n d a r d d e n s i t y a n d t h e O D v a l u e , w i t h t h e s a m p l e O D v a l u e i n t h e e q u a t i o n , c a l c u l a t e t h e s a m p l e d e n s i t y , m u l t i p l i e d b y t h e d i l u t i o n m u l t i p l e , t h e r e s u l t i s t h e s a m p l e a c t u a l d e n s i t y .
I m p o r t a n t n o t e s
T h e k i t t a k e s o u t f r o m t h e r e f r i g e r a t i o n e n v i r o n m e n t s h o u l d b e b a l a n c e d 1 5 - 3 0 m i n u t e s i n t h e r o o m t e m p e r a t u r e t h e n u s e , E L I S A p l a t e s c o a t e d i f h a s n o t u s e u p a f t e r o p e n e d , t h e p l a t e s h o u l d b e s t o r e d i n S e a l e d b a g .
w a s h i n g b u f f e r w i l l C r y s t a l l i z a t i o n s e p a r a t i o n , i t c a n b e h e a t e d t h e w a t e r h e l p s d i s s o l v e w h e n d i l u t e . W a s h i n g d o e s n o t a f f e c t t h e r e s u l t .
a d d S a m p l e w i t h s a m p l e r E a c h s t e p , A n d p r o o f r e a d i t s a c c u r a c y f r e q u e n t l y , a v o i d s t h e e x p e r i m e n t a l e r r o r . a d d s a m p l e w i t h i n 5 m i n , i f t h e n u m b e r o f s a m p l e i s m u c h , r e c o m m e n d t o u s e V o l l e y .
P l e a s e m a k e s p e c i f i c a t i o n c u r v e w h e n y o u a s s a y , h a d b e t t e r m a k e d u p l i c a t e w e l l , i f i n t h e s a m p l e t h e t e s t i n g m a t e r i a l c o n t e n t i s e x c e s s i v e l y h i g h ( T h e s a m p l e O D i s b i g g e r t h a n t h e f i r s t s t a n d a r d w e l l O D ) , p l e a s e u s e S a m p l e d i l u t i o n t o d i l u t e c e r t a i n m u l t i p l e ( n t i m e s ) , t h e n a s s a y . P l e a s e m u l t i p l y t o t a l D i l u t i o n T i m e s w h e n c a l c u l a t e n 5 .
P l e a s e a c c o r d i n g t o u s e i n s t r u c t i o n s t r i c t l y , T h e t e s t r e s u l t d e t e r m i n a t i o n m u s t t a k e t h e m i c r o t i t e r p l a t e r e a d e r a s a s t a n d a r d .
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