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 ƒ  ½ ¨ 7 Hë  H  Sae Mulli (The Korean Physical Society), Volume 49, Number 5, 2004¸   11 Z 4, pp. 418∼421

NiCl 2   Â Ê S Ëc Ü R ‰ ˜ ms œ ù m Ç ú m É M “ ˜ m8 ý < g ºU ê s0 n É; c 6 ” X ¢ R w ‹ Ž ì ŏ Œ

g `

@+ Ö <4 w H · * × <Q * å 

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–² D G @ /† < Ɠ §  ƒ  õ † < ÆÂ Ò 6 £ x6   xÓ ü t o † < Ƅ  / B N, " fÖ  ¦ 140-714 (2004¸   9 Z 4 21{ 9  ~ à Î6 £ §)

é ß

–Ñ þ ˜| 9  } 9  ] j› ¸\  e ” # Q m Ö 0 qs   ïh A ) a Ä »o  l ó ø ÍÜ ¼– Ð Histidine-tagged é ß –Ñ þ ˜| 9 `  ¦ “ ¦& ñ  or v   H  Œ •

\ O

“ É r B Ä º ×  æ כ ¹ô  Ç { 9 s  . ‘ : r z  ´+ « >\ " f  H Histidine-tagged é ß –Ñ þ ˜| 9 _  “ ¦& ñ  o\  ¦ 0 AK  Û ¼— 2 ; ïh AZ O õ  n  i ç

Z O \  _ K  NiCl

2

\  ¦ Ä »o l ó ø Í 0 A\   ïh A % i  . s X O >  ] j› ¸  ) a l ó ø Í 0 A\  fluorescein isothiocyanate (FITC)-conjugated é ß –Ñ þ ˜| 9 õ  < ʓ É r FITC conjugated ÷ &# Q e ” Ü ¼€  " f 1 l x r \  Histidine-tagged s  · ¡ ­ # Q e ”

  H WAX-9D é ß –Ñ þ ˜| 9 `  ¦ ì ø Í6 £ x r &  Ä »o  l ó ø Í\  “ ¦& ñ  oô  Ç Ê ê Bio-imaging Analyzer System`  ¦ s 6   x 

#

Œ é ß –Ñ þ ˜| 9   ҂ à Ì_ …Û ¼à Ô\  ¦ z  ´' Ÿ  % i  . m Ö 0 q  ïh A l ó ø Í_  ³ ð€   : £ ¤$ í “ É r Particle Size Analyzer ü < AFM

`

 ¦  6   x # Œ ì  r$ 3  % i  . ‘ : r z  ´+ « >\ " f Û ¼— 2 ; ïh AZ O “ É r Ä »o  l ó ø ÍÜ ¼– Ð á Ԗ Ж 2 ; } 9  ] j› ¸\  e ” # Q a % ~“ É r — ¸ 4

S qs  | ¨ c  כ Ü ¼– Ð ¨ î   ) a  .

PACS numbers: 68, 62, 81, 87

Keywords: é ß –Ñ þ ˜| 9 } 9  e  ¦ Y Us à Ô, a % ¦- 0 q ~ ½ ÓZ O , n i ç ~ ½ ÓZ O , y Û ¼I Õ ª, % i  om Ö 0 q

I. " e  ] Ø

é

ß –Ñ þ ˜| 9 ^ ‰_  4 Ÿ ¤½ + Ë& h “   l 0 p x ì  r$ 3 “ É r þ j  H Ä »„  ^ ‰\  @ / ô 

Ç ƒ  ½ ¨ “  ç ß – DNA % i l " f\ P `  ¦ ¢ - a„  y  µ 1 ß) €? /€  " f  Ö ¸ µ

1 Ï >  ”  ' Ÿ ÷ &  H  כ õ   8Ô  ¦ # Q Ò q t" î ‰ & ³ © œõ  | 9 # î _  l " é ¶

`

 ¦ s K    H X < € 9 à º& h Ü ¼– Ð כ ¹½ ¨÷ &# Q”   . “  ç ß – Ä »„  ^ ‰ _

 ƒ  ½ ¨  õ \   Ø Ô€   “  ç ß –“ É r 2ë ß – ∼ 3ë ß – > h_  Ä »„   

\

 ¦ t “ ¦ e ” Ü ¼ 9, s  ´ ú §“ É r Ä »„   _  1 l x r  µ 1 Ï& h  ƒ  ½ ¨

\

 ¦ 0 AK   s ß ¼– Ð# QY Us (microarray) Ä »„    } 9 `  ¦ s 6   x

“ ¦ e ”   [1–4].  s ß ¼– Ð# QY Us  Ä »„    } 9 “ É r à ºÑ þ ˜ ∼ à º … 

; > h_  Ä »„   \  ¦ p [ jô  Ç / B Nç ß – 0 A\  “ ¦& ñ  or &  1 l x r   µ

1 Ï& h Ü ¼– Ð Ã ºÑ þ ˜ ∼ à ºë ß – > h_  Ä »„   _     o\  ¦ Ä »„   ç ß – _

  © œ˜ Ð& h    ½ + Ë`  ¦ : Ÿ x # Œ & ñ | ¾ Óì  r$ 3  † < ÊÜ ¼– Ð+ ‹ Ä »„    µ 1 Ï

‰ &

³s   [  tƒ    s   Ž Ò  o\  s 6   x ÷ &“ ¦ e ”  . é ß –Ñ þ ˜| 9 “ É r Ä »„  



 µ 1 ω & ³`  ¦ : Ÿ x K  ë ß –[ þ t # Qt   H “ ¦ì  r   Ó ü t| 9 – Ð ô  Ç Ä »„   – Ð Â

Ò'  # Œ Q > h_  é ß –Ñ þ ˜| 9 – Ð Ò q tí ß – 0 p xô  Ç  כ \   8 # Œ, „   s

 Ê ê    o (posttranslational modification)– Ð “  K  Ä »„  



˜ Ð   s `›    € ª œô  Ç 7 á x À Ó e ”  . 7 £ ¤, “  ç ß –s  ° ú “ ¦ e ”   H é

ß –Ñ þ ˜| 9 _  7 á x À Ӎ  H Ä »„   _  t  à º\  ¦  s `›   0 p x ô  Ç “ ¦

^

 ¦ à º e ”  .   " f é ß –Ñ þ ˜| 9 ^ ‰_  1 l x r  µ 1 Ï& h  ì  r$ 3 “ É r Ä »„  

^

‰_  ƒ  ½ ¨˜ Ð  † ¾ ÓÊ ê Ò q t" î ‰ & ³ © œõ  | 9 # î ƒ  ½ ¨\  Ä »„  ^ ‰_ 

ƒ 

½ ¨˜ Ð   8¹ ¡ ¤ € 9 à º& h s   ½ + É Ã º e ”  . é ß –Ñ þ ˜| 9 ^ ‰_  1 l x r 

E-mail: [email protected];

Tel: +82-2-709-2813; Fax: +82-2-792-6082



µ 1 Ï& h  ì  r$ 3 `  ¦ 0 Aô  Ç ƒ  ½ ¨~ ½ ÓZ O  ×  æ   Ä »„   _  1 l x r 



µ 1 Ï& h  ƒ  ½ ¨~ ½ ÓZ O _   “   DNA } 9  ì  r$ 3 ~ ½ ÓZ O % ƒ! 3  é ß –Ñ þ ˜

| 9

} 9 `  ¦ s 6   xô  Ç ì  r$ 3 ~ ½ ÓZ O s   [5]. é ß –Ñ þ ˜| 9 } 9  ] j› ¸_   © œ l

œ í& h  l Õ ü t“ É r é ß –Ñ þ ˜| 9 `  ¦ Ä »o  l ó ø Í 0 A\   ҂ à Ìr v l  0 A ô 

Ç l ó ø Í_  „  % ƒo  l Õ ü t – Ð s  כ “ É r é ß –Ñ þ ˜| 9 _  l ó ø Í Â Ò‚ à Ì~ ½ Ó Z O

õ  & h ½ + Ë >  s À Ò# Q4 R ô  Ç  [5–7].

‘

: r ƒ  ½ ¨\ " f  H é ß –Ñ þ ˜| 9  ³ ð€   e ” _ _   Òì  r s  l ó ø Í 0 A\  Â

҂ à Ì÷ &  H  כ `  ¦ ~ ½ Ót  l  0 A # Œ Ni

2+

s “ : r \  : £ ¤ s & h Ü ¼– Ð

 

½ + Ë   H Histidine-tagged ÷ &# Q e ”   H é ß –Ñ þ ˜| 9 `  ¦ l ó ø Í 0 A

\

   ½ + Ër ~  ´ 3 l q& h Ü ¼– Ð m Ö 0 qs “ : r s   ҂ à ̝ ) a l ó ø Í`  ¦ ] j› ¸

% i  .   " f t F K  t  ˜ Г ¦  ) a é ß –Ñ þ ˜} 9 } 9  Ä »o  l ó ø Íõ 

°

ú  s     l ü < · ú ˜X <y × ¼l  1 p x s  l ó ø Í³ ð€  \  ” ¸Ø  ¦ ÷ &• ¸2 Ÿ ¤ n

i ç (dipping) ~ ½ ÓZ O õ  Û ¼— 2 ; ïh A(spin coating) ~ ½ ÓZ O \   



 é ß –Ñ þ ˜| 9 } 9  l ó ø Í`  ¦ ] j› ¸ô  Ç Ê ê, l ó ø Í ³ ð€    © œI \  ¦ q “ § ì

 r$ 3  “ ¦ é ß –Ñ þ ˜| 9  “ ¦& ñ  o  Œ •\ O `  ¦ à º' Ÿ  % i  . Õ ª1 l xî ß – é ß – Ñ þ

˜| 9 } 9  ] j› ¸ l Õ ü t“ É r ç ß –¼ # $ í õ   â ] j$ í _  8 £ ¤€  \ " f n i ç

~

½ ÓZ O s  Å Ò\  ¦ s À Ò% 3 Ü ¼  l ó ø Í_  ³ ð€   > h| 9  8 £ ¤€  \ " f Û ¼

—

2 ;  ïh A ~ ½ ÓZ O `  ¦ & h 6   x # Œ s  ¿ º t  ~ ½ ÓZ O `  ¦ q “ § ì  r$ 3  ô 

Ç \ V  H  f ”  ë  H‰  ³\  ˜ Г ¦  ) a   \ O Ü ¼ 9, ‘ : r ƒ  ½ ¨  H s 

\

 › ' aô  Ç ' Í ˜ Г ¦   H €  \ " f ×  æ כ ¹ô  Ç _ p \  ¦ t    “ ¦ ½ + É Ã

º e ”  ’ x .

II. ÷ m Ç] M öU ê s0 n É

-418-

(2)

 ƒ  ½ ¨ 7 Hë  H  NiCl

2

  ïh A ) a é ß –Ñ þ ˜| 9  } 9  l ó ø Í_  ] j› ¸~ ½ ÓZ O \  @ /ô  Ç q “ § ƒ  ½ ¨ – ‰ & ³ï  r" é ¶ · î  r p % ò -419-

Fig. 1. Photos of nickel chloride coated plates. Left panel : by dipping method, Right panel : by spin-coating method.

1. M “ ˜ m8 ý < g º

Ä

»o  l ó ø Í 0 A\  NiCl

2

} Œ •`  ¦ n i ç ~ ½ ÓZ O õ  Û ¼— 2 ;  ïh A

~

½ ÓZ O Ü ¼– Ð + þ A$ í r (   . 6   xÓ  o ½ + Ë$ í `  ¦ 0 Aô  Ç " é ¶ « ÑÓ ü t| 9 – Ѝ  H NiCl

2

·6H

2

O ü < 6   x B – Ѝ  H 7 £ x À Óà º\  ¦  6   x % i  . ] j› ¸  ) a 6

 

xÓ  o`  ¦ Ä »o  l ó ø ÍÜ ¼– Ð  ïh A l  „  \  Ä »o  l ó ø Í 0 A_  Ô  ¦ í

 HÓ ü t ] j \  ¦ 0 AK  \ ò ø Í`  ¦ õ   [ j— : r`  ¦  6   x # Œ 15 ì  r 1 l x î

ß – œ í6 £ §  [ j' ‘  Ê ê | › ¸r (   . NiCl

2

} Œ •`  ¦ ] j› ¸ l  0 A K

 2500 rpm_  Û ¼— 2 ; ï' \  ¦  6   x % i Ü ¼ 9, } Œ • ] j› ¸ Ê ê\ 



 H ³ ð€   î ß –& ñ  o\  ¦ 0 A # Œ 80

C _  í ß –™ è ì  r0 Al \ " f 5ì  r 1

l

xî ß – \ P % ƒo  % i  . ] j› ¸  ) a NiCl

2

} Œ •_  ³ ð€   : £ ¤$ í `  ¦ › ' a¹ 1 Ï

l  0 AK  PSA(Particle size analyzer, IMT i-30) ì  r$ 3 õ  AFM ì  r$ 3 `  ¦ à º' Ÿ  % i  .

2. ‰ ˜ ms œ ù m Ç w ŠX N Ë× D

é

ß –Ñ þ ˜| 9  “ ¦& ñ  o z  ´+ « >“ É r NiCl

2

s   ïh A ) a Ä »o l ó ø Í 0 A

\

 fluorescein isothiocyanate (FITC)-conjugated ÷ &# Q e ”

Ü ¼€  " f 1 l x r \  Histidine-tagged s  · ¡ ­ # Q e ”   H WAX- 9D é ß –Ñ þ ˜| 9 `  ¦ 10 µg/ml 0 l x • ¸– Ð 10 ì  r \ " f ô  Ç r ç ß – 1 l xî ß – z 

´“ : r \ " f ì ø Í6 £ x r †   + ' phosphate buffered saline(PBS)

¢ -

aØ  æÓ  oÜ ¼– Ð 5ì  rç ß – [ j    [ j' ‘  # Œ | › ¸ô  Ç Ê ê + þ AF g image analyzer – Ð ì  r$ 3  % i  . Ä »o  l ó ø Í\  “ ¦& ñ  oô  Ç FITC-conjugated é ß –Ñ þ ˜| 9 “ É r Bio-Imaging Analyzer Sys- tem (BAS; Fuji Photo Film Co) \ " f excitation  © œ 490 nm ü < emission  © œ 670 nm– Ð “ ¦& ñ # Œ 8 £ ¤& ñ % i  .

III. + s ÇÊ Ý õ m Í w в  o

Û

¼— 2 ; ïh A ~ ½ ÓZ O “ É r F K5 Å q_  Ä »l  x 9 Á ºl  o½ + ËÓ ü t_  6   xÓ  o

`

 ¦ l ó ø Í\  • ¸Ÿ íô  Ç Ê ê B Ä º   É r  r„  à º– Ð l ó ø Í`  ¦  r„  r 

&

 " é ¶   H Ó ü t| 9 `  ¦ l ó ø Í ³ ð€  \   ïh Ar v   H ~ ½ ÓZ O Ü ¼– Ð ç  H

Fig. 2. Particle size analysis for the glass slides coated with there different NiCl

2

concentrations. The nickel chloride-coated slides were prepared as described in the results and discussion and the analyzed by PSA(5,000).

Left panel : by spin-coating method, Right panel : by dipping method.

{ 9

ô  Ç l ó ø Í} Œ •`  ¦ + þ A$ í r ~  ´ à º e ” `  ¦ ÷  rë ß –  m   { 9 ì ø Í& h “  



ïh A ~ ½ ÓZ O \  q K   ïh A5 Å q • ¸ B Ä º   É r  © œ& h `  ¦ t    .

ì

ø ̀  \  n i ç ~ ½ ÓZ O “ É r l ó ø Í`  ¦  ïh A “ ¦    H 6   xÓ  o 5 Å q \  f ”

] X  V , % 3   N S  H é ß –í  Hô  Ç ~ ½ ÓZ O Ü ¼– Ð  ïh A / B N& ñ s  B Ä º é

ß –í  H t ë ß – l ó ø Í ³ ð€  _   ïh A  © œI   H ç  H{ 9 $ í `  ¦ ° ú t  3 l w

  H é ß –& h s  e ”  . Ä º‚   ] j› ¸  ) a l ó ø Í} Œ •_  : £ ¤$ í `  ¦ › ¸   l

 0 Aô  Ç 8 £ ¤& ñ [ þ t`  ¦ à º' Ÿ  % i  . Fig. 1 (a)  H n i ç ~ ½ ÓZ O \  _ K , (b)  H Û ¼— 2 ;  ïh A ~ ½ ÓZ O \  _ K  ] j› ¸  ) a l ó ø Í_  z  ´] j

—

¸_ þ v s  . Õ ªa Ë >\ " f ˜ Ѝ  H  ü < ° ú  s  n i ç ~ ½ ÓZ O \  _ K  ] j

›

¸  ) a l ó ø Í[ þ t_  ³ ð€  “ É r B Ä º  } 9 “ ¦ ™ D ¥„ à Ìô  Ç  © œI \  ¦ ˜ Ð# Œ Å

ҍ  H ì ø ̀   Û ¼— 2 ;  ïh A ~ ½ ÓZ O \  _ K  ] j› ¸  ) a l ó ø Í[ þ t_  ³ ð€  

“ É

r q “ §& h  B ã ¼X O “ ¦ L :  F Mô  Ç  © œI – Ð › ' a¹ 1 Ï÷ &% 3  . Ó ü t : r n  i ç

~ ½ ÓZ O \  _ K  ] j› ¸  ) a l ó ø Í_   â Ä º } Œ • ¿ ºa \  ¦ 7 £ x r ~  ´ Ã

º  H e ” Ü ¼  ³ ð€  _  „  ì ø Í& h “   NiCl

2

} Œ •_  ç  H{ 9 $ í “ É r ß ¼>  b 

# Qt   H  כ Ü ¼– Ð ¨ î   ) a  . Õ ª Q  Û ¼— 2 ;  ïh A ~ ½ ÓZ O \  _  K

 ] j› ¸  ) a l ó ø Í“ É r } Œ •_  ç  H{ 9 $ í `  ¦ ß ¼>  7 £ x r v   H 1 l x r 

\

 é ß –Ñ þ ˜| 9 } 9  l ó ø ÍÜ ¼– Ð" f_   © œ¾ ¡ §$ í `  ¦ ° ú Ø  ¦ à º e ” `  ¦  כ Ü ¼

–

Ð l @ /  ) a  . 7 £ ¤,  ïh A ³ ð€  s  ç  H{ 9 ½ + É  â Ä º é ß –Ñ þ ˜| 9   Ö ¸$ í Â

Ò0 A Ä »o  l ó ø Í „  €  \    5 g { 9 & ñ >  ” ¸Ø  ¦ ÷ &# Q é ß –Ñ þ ˜

| 9

_   ҂ à ̧ 4 `  ¦ 7 £ x r v >   ) a  . Fig. 2  H Particle size analyzer _    õ \  ¦ ˜ Ð# Œï  r  . Fig. 2_  (a), (b), (c)  H Û ¼

—

2 ;  ïh A ~ ½ ÓZ O Ü ¼– Ð 7 £ x À Óà ºü < NiCl

2

_  2:1, 10:1, 50:1 0 l x

•

¸q – Ð ] j› ¸  ) a l ó ø Í_  ³ ð€   + þ A © œ`  ¦    · p . 7 £ x À Óà ºü <

(3)

-420- ô  Dz D GÓ ü t o † < Æ rt  “D hÓ ü t o ”, Volume 49, Number 5, 2004¸   11 Z 4

NiCl

2

_  0 l x • ¸q  y Œ ™™ è† < Ê\     ³ ð€  _  Ni

2+

s “ : r_  € ª œ s

 y Œ ™™ è† < ÊÜ ¼– Ð+ ‹ é ß –Ñ þ ˜| 9 _   ҂ Ã Ì 0 p x§ 4 `  ¦ y Œ ™™ èr v   H f ”  ] X

& h “   " é ¶ “  s   ) a  . s  Qô  Ç   õ   H NiCl

2

ì  r   { © œ é ß –Ñ þ ˜

| 9

  ҂ Ã Ì  Ö ¸$ í  Ò0 A ô  Ç > hm ”  e ”   H X <, 0 l x • ¸ y Œ ™™ è– Ð “  K 

 Ö

¸$ í  Ò0 A > hà º y Œ ™™ èÙ þ ¡l  M :ë  H“    כ Ü ¼– Ð ˜ Ð# Œ ”   .

Fig. 2_  (d), (e), (f)  H n i ç ~ ½ ÓZ O Ü ¼– Ð 7 £ x À Óà ºü < NiCl

2

_  2:1, 10:1, 50:1 0 l x • ¸q – Ð ] j› ¸  ) a l ó ø Í_  ³ ð€   + þ A © œ`  ¦  

 · p . s  % i r  0 l x • ¸ y Œ ™™ è– Ð “  K  l ó ø Í ³ ð€  _   Ö ¸$ í Â Ò 0 A y Œ ™™ è÷ &# Q é ß –Ñ þ ˜| 9 _   ҂ Ã Ì 0 p x§ 4 `  ¦ b  # Qä ¼o >  ÷ & 9, Û

¼— 2 ;  ïh A ~ ½ ÓZ O Ü ¼– Ð ] j› ¸  ) a l ó ø Í_  ³ ð€  õ  ² ú ˜o  { 9  [ þ t s

 ß ¼>  é # Qo t “ ¦ Ô  ¦ ½ ©g Ë :& h “   — ¸_ þ v`  ¦ ˜ Ð# Œ ï  r  . Fig.

3(a), (b)  H Û ¼— 2 ;  ïh A ~ ½ ÓZ O \  _ K , Fig. 3(c), (d)  H n i ç

~

½ ÓZ O \  _ K  7 £ x À Óà ºü < NiCl

2

_  2:1 õ  50:1 0 l x • ¸q – Ð ] j

›

¸  ) a l ó ø Í_  AFM % ò  © œ`  ¦ ˜ Ð# Œ ï  r  . Õ ªa Ë >\ " f ˜ Ѝ  H   ü

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–

Ð › ' a¹ 1 Ï÷ &% 3  . Õ ª Q  Fig. 3(c)_  n i ç ~ ½ ÓZ O Ü ¼– Ð ] j› ¸

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a l ó ø Í_   â Ä º   É r l ó ø Í_  ³ ð€  \  q K  NiCl

2

} Œ •s  ¿ º , 

>   ïh A÷ &% 3 Ü ¼ , Fig. 4\ " f ˜ Ѝ  H  ü < ° ú  s  é ß –Ñ þ ˜| 9 

“

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 Ä »o  l ó ø Í\  y © œ >    ½ + Ë÷ &t  3

l

w† < ÊÜ ¼– Ð+ ‹ é ß –Ñ þ ˜| 9 `  ¦  ҂ à Ìr †   Ê ê 0 >d ç (washing)  Œ •\ O  r

 NiCl

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 ~ 1 >  b  # Q4 R    H  כ Ü ¼– Ð ˜ Ð# Œ ”   . Fig.

4  H Û ¼— 2 ;  ïh A ~ ½ ÓZ O õ  n i ç ~ ½ ÓZ O Ü ¼– Ð 7 £ x À Óà ºü < NiCl

2

_  0

l

x • ¸    o\    É r l ó ø Í  © œ_  é ß –Ñ þ ˜| 9   ҂ Ã Ì 0 p x§ 4 `  ¦ ˜ Ð# Œ ï

 r  . Histidine-tagged é ß –Ñ þ ˜| 9 “ É r Ä »„    › ¸½ + Ë`  ¦ s 6   x 

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Œ é ß –Ñ þ ˜| 9 \  6> h_  histidine`  ¦ ƒ  5 Å q& h Ü ¼– Ð ³ ðt ô  Ç  כ Ü ¼

–

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3  . s   H Fig. 3 \ " f [ O " î ô  Ç  ü < ° ú  s  Û ¼— 2 ;  ïh A ~ ½ ÓZ O  Ü

¼– Ð ] j› ¸  ) a l ó ø Í_   â Ä º NiCl

2

 Ä »o  ³ ð€  õ  ˜ Ð  x 9  ] X

 >    ½ + Ë| ¨ c à º e ”   H ¨ 8 Š â `  ¦ ] j/ B Nô  Ç  כ Ü ¼– Ð  « Ñ  ) a  .



 " f Û ¼— 2 ;  ïh A ~ ½ ÓZ O Ü ¼– Ð ] j› ¸  ) a m Ö 0 q  ïh A l ó ø Í“ É r l  ó

ø Í 0 A\   ҂ à Ì÷ &  H His-tag  ) a  ¿ é ß –Ñ þ ˜| 9 _  l 0 p x ’ < H © œs  n

i ç ~ ½ ÓZ O \  q K  B Ä º ± ú l  M :ë  H \  é ß –Ñ þ ˜| 9  } 9 6   x l ó ø ÍÜ ¼

–

Ð B Ä º & h ½ + ˽ + É  כ Ü ¼– Ð ¨ î   ) a  .

IV. + s Ç Â ] Ø

‘

: r ƒ  ½ ¨\ " f  H NiCl

2

  ïh A ) a é ß –Ñ þ ˜| 9  } 9  l ó ø Í ] j› ¸

\

 ¦ 0 AK  Û ¼— 2 ;  ïh A ~ ½ ÓZ O õ  n i ç ~ ½ ÓZ O `  ¦ q “ § ì  r$ 3  % i 

Fig. 3. AFM micrograph for the glass slides coated with various concentrations of NiCl

2

. The nickel chloride- coated slides were prepared as described in the Discus- sion and Results and then analyzed by AFM. (a) and (b)

; 50 % and 2 % NiCl

2

by spin coating method, (c) and (d) ; 50 % and 2 % NiCl

2

by dipping method.



. 7 £ x À Óà ºü < NiCl

2

_  0 l x • ¸    o\     l ó ø Í_  : £ ¤$ í `  ¦

“

¦¹ 1 Ïô  Ç   õ , Û ¼— 2 ;  ïh A ~ ½ ÓZ O Ü ¼– Ð ] j› ¸ô  Ç l ó ø Í_  ³ ð€  : £ ¤

$ í

õ  é ß –Ñ þ ˜| 9   ҂ Ã Ì 0 p x§ 4 s  Ä ºÃ ºô  Ç  כ Ü ¼– Ð ¨ î ÷ &% 3  . 7 £ x À

Óà ºü < NiCl

2

_  0 l x • ¸q  y Œ ™™ è† < Ê\     ³ ð€  _  Ni

2+

s 

“

: r_  € ª œs  y Œ ™™ è† < ÊÜ ¼– Ð+ ‹ é ß –Ñ þ ˜| 9 _   ҂ Ã Ì 0 p x§ 4 `  ¦ y Œ ™™ èr  v

  H f ” ] X & h “   " é ¶ “  s  ÷ &  H X <, s  Qô  Ç   õ   H NiCl

2

ì  r   {

© œ é ß –Ñ þ ˜| 9   ҂ Ã Ì  Ö ¸$ í  Ò0 A_  > hà º y Œ ™™ èÙ þ ¡l  M :ë  H“    כ Ü

¼– Ð ˜ Ð# Œ ”   . : £ ¤ y  n i ç ~ ½ ÓZ O Ü ¼– Ð ] j› ¸  ) a l ó ø Í_   â Ä

º NiCl

2

} Œ •s  ¿ º,  >   ïh A÷ &% 3 Ü ¼  é ß –Ñ þ ˜| 9  “ ¦& ñ  o 0 p x§ 4 

“ É

r b  # Qt   H  כ Ü ¼– Ð ¨ î   ) a  . s   H Û ¼— 2 ;  ïh A ~ ½ ÓZ O \  q 

Fig. 4. The His-tagged protein was specially adsorbed

to the NiCl

2

coated glass slides. The nickel coated slides

were constructed by (a) the spin-coating method and (b)

the dipping method using the indicated amount of NiCl

2

.

(4)

 ƒ  ½ ¨ 7 Hë  H  NiCl

2

  ïh A ) a é ß –Ñ þ ˜| 9  } 9  l ó ø Í_  ] j› ¸~ ½ ÓZ O \  @ /ô  Ç q “ § ƒ  ½ ¨ – ‰ & ³ï  r" é ¶ · î  r p % ò -421-

K

 NiCl

2

 Ä »o  l ó ø Í\  y © œ >    ½ + Ë÷ &t  3 l w† < ÊÜ ¼– Ð+ ‹ é ß – Ñ þ

˜| 9 `  ¦  ҂ à Ìr †   Ê ê 0 >d ç (washing)  Œ •\ O  r  NiCl

2

 ~ 1 

>

 b  # Q4 R    H  כ Ü ¼– Ð ˜ Ð# Œ ”   . s  כ “ É r AFM ì  r$ 3  õ

 é ß –Ñ þ ˜| 9  “ ¦& ñ  o z  ´+ « >\ " f S X ‰ “  ÷ &% 3  .

P c

p 8 ý ò k >

s

 ƒ  ½ ¨  H 2004† < Ƹ  • ¸ é ß –² D G @ /† < Ɠ § @ /† < ƃ  ½ ¨q _  t " é ¶ Ü

¼– Ð ƒ  ½ ¨÷ &% 3 _ þ v m  .

Y c

p w Š à U Ø ”  ô

[1] H. Yasuda and C. E. Lamage, J. Appl. Polym. Sci.

17, 201 (1973).

[2] H. Muguruma and I. Karube, Trends Anal. Chem.

18, 62 (1999).

[3] K. Naranishi, H. Muguruma and I. Karube, Anal.

Chem. 68, 68 (1996).

[4] H. Miyachi, A. Hiratsuka, K. Ikebukuro, K. Yano, H.

Muguruma and I. Karube, Biotechnol. Bioengin. 69, 323 (2000).

[5] G. MacBeath and S. L. Schreiber, Science 289, 1760 (2000).

[6] D. V. Nicolau, H. Taguchi, H. Taniguchi and S.

Yoshikawa, Micron-sized Protein Patterning on Di- azonaphthoquinone/novolak Thin Polymaric Films.

Langmuir 14, 1927 (1998).

[7] R. Bashir, R. Gomez, A. Sarikaya, M. R. Ladisch, J.

Sturgis and J. P. Robinson, Adsorption of Avidin on Microfabricated Surface for Protein Biochip Applica- tions, Biotechnol. Bioengin. 73, 324 (2001).

A Comparative Study for the Fabrication of Nickel Chloride Coated Protein Chip Plates

June Won Hyun

and Mi Young Yun

Applied Physics, College of Natural Science, Dankook University, Seoul 140-714 (Received 21 September 2004)

The adsorption of proteins on the surfaces of glass slides is essential for the construction of protein chips. Here, we report that histidine-tagged proteins can be efficiently adsorbed on glass slides coated with nickel. A variety of nickel chloride-coated plates are prepared by using either a spin-coating or a dipping method, and experiments on the adsorption of histidine-tagged proteins were carried out on these plates. The extent of the protein adsorption was determined by using a bio-imaging analyzer system after the slides ahd been incubated with a cross-linker, 1-ethyl-3-[3- dimethylaminopropyl] carbodi-imide hydrochloride and a fluorescein isothiocyanate-tagged WAX- 9D protein. The properties of the nickel chloride thin layer on the surface of the slides were assayed using a particle size analyzer and the surfaces status of the coated slides was examined by using an atomic force microscope. The spin-coating method presented here is shown to be a promising technique for coat glass slides for the fabrication of protein chips.

PACS numbers: 68, 62, 81, 87

Keywords: Protein chip plate, Sol-gel method, Dipping method, His-tagged, Nickel chloride

E-mail: [email protected]

수치

Fig. 1. Photos of nickel chloride coated plates. Left panel : by dipping method, Right panel : by spin-coating method
Fig. 3. AFM micrograph for the glass slides coated with various concentrations of NiCl 2

참조

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