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Spin columnŸÀÔÀ¸·Î ºü¸£°í °£ÆíÇÑ His-tag ´Ü¹éÁú Á¤Á¦

Capturem¢â His-Tagged Purification

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Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
ºñ°í »ç¿ëÀڸŴº¾ó
Clontech
635710
Capturem¢â His-Tagged Purification Miniprep Kit
°ü·ÃÇмú °ü½É»óǰµî·Ï ±¸¸ÅÇϱ⠶óÀ̼±½º 
20ȸ
296,800¿ø 
371,000¿ø
°¡°ÝÇÒÀÎ
11.01 ~ 12.13
Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
Capturem His-Tagged Purification Miniprep Kit Protocol-At-A-Glance_042716.pdf, xTractor Buffer Baculovirus Sample Preparation Protocol-At-A-Glance_031616.pdf, xTractor%20Buffer%20and%20xTractor%20Buffer%20Kit%20User%20Manual%20(PT3735-1)_031616.pdfxTractor%20Buffer%20Bacterial%20Sample%20Preparation%20Protocol-At-A-Glance%20(PT3735-2)_031716.pdf, xTractor%20Buffer%20Mammalian%20Sample%20Preparation%20Protocol-At-A-Glance_031616.pdf
Clontech
635713
Capturem¢â His-Tagged Purification Maxiprep Kit
°ü·ÃÇмú °ü½É»óǰµî·Ï ±¸¸ÅÇϱ⠶óÀ̼±½º 
6 ȸ
386,400¿ø 
483,000¿ø
°¡°ÝÇÒÀÎ
11.01 ~ 12.13
Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
xTractor Buffer Mammalian Sample Preparation Protocol-At-A-Glance_031616.pdf, xTractor Buffer Bacterial Sample Preparation Protocol-At-A-Glance (PT3735-2)_031716.pdf, Capturem%20His-Tagged%20Purification%20Miniprep%20Kit%20Protocol-At-A-Glance_121918.pdf, xTractor%20Buffer%20and%20xTractor%20Buffer%20Kit%20User%20Manual%20(PT3735-1)_031616.pdf, xTractor%20Buffer%20Baculovirus%20Sample%20Preparation%20Protocol-At-A-Glance_031616.pdf, Capturem%20His-Tagged%20Purification%20Maxiprep%20Kit%20Protocol-At-A-Glance_121918.pdf
Clontech
635719
Capturem¢â His-Tagged Purification Maxiprep Columns
°ü·ÃÇмú °ü½É»óǰµî·Ï ±¸¸ÅÇϱ⠶óÀ̼±½º 
50°³
1,524,000¿ø 
1,905,000¿ø
°¡°ÝÇÒÀÎ
11.01 ~ 12.13
Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
Capturem His-Tagged Purification Maxiprep Columns Protocol-At-A-Glance_042716.pdf, xTractor Buffer Bacterial Sample Preparation Protocol-At-A-Glance (PT3735-2)_031716.pdf, xTractor%20Buffer%20and%20xTractor%20Buffer%20Kit%20User%20Manual%20(PT3735-1)_031616.pdf, xTractor%20Buffer%20Baculovirus%20Sample%20Preparation%20Protocol-At-A-Glance_031616.pdf, xTractor%20Buffer%20Mammalian%20Sample%20Preparation%20Protocol-At-A-Glance_031616.pdf, xTractor%20Buffer%20Yeast%20Sample%20Preparation%20Protocol-At-A-Glance_031616.pdf
Clontech
635724
Capturem¢â His-Tagged Purification Large Volume
°ü·ÃÇмú °ü½É»óǰµî·Ï ±¸¸ÅÇϱ⠶óÀ̼±½º 
4 ȸ
636,800¿ø 
796,000¿ø
°¡°ÝÇÒÀÎ
11.01 ~ 12.13
Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
Capturem His-Tagged Purification Large Volume Protocol-At-A-Glance_061317.pdf
Clontech
635714
Capturem¢â His-Tagged Purification 96
°ü·ÃÇмú °ü½É»óǰµî·Ï ±¸¸ÅÇϱ⠶óÀ̼±½º 
1 plate
547,200¿ø 
684,000¿ø
°¡°ÝÇÒÀÎ
11.01 ~ 12.13
Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
Capturem His-Tagged Purification 96 Protocol-At-A-Glance_042716.pdf, xTractor Buffer Baculovirus Sample Preparation Protocol-At-A-Glance_031616.pdf, Capturem%20His-Tagged%20Purification%20Miniprep%20Kit%20Protocol-At-A-Glance_121918.pdf, xTractor%20Buffer%20and%20xTractor%20Buffer%20Kit%20User%20Manual%20(PT3735-1)_031616.pdf, xTractor%20Buffer%20Bacterial%20Sample%20Preparation%20Protocol-At-A-Glance%20(PT3735-2)_031716.pdf, xTractor%20Buffer%20Mammalian%20Sample%20Preparation%20Protocol-At-A-Glance_031616.pdf
Clontech
635730
Capturem¢â His-Tagged Purification 24-Well Plate
°ü·ÃÇмú °ü½É»óǰµî·Ï ±¸¸ÅÇϱ⠶óÀ̼±½º 
1 plate
559,200¿ø 
699,000¿ø
°¡°ÝÇÒÀÎ
11.01 ~ 12.13
Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â
Capturem His-Tagged Purification 24-Well Plate Protocol-At-A-Glance_040518.pdf

Tip. ´õ¿í È¿À²ÀûÀÎ ´Ü¹éÁú ÃßÃâ°ú Á¤Á¦¸¦ À§ÇÏ¿© xTractor Buffer ¸Å´º¾óÀÇ Lysis protocol(Ŭ¸¯)À» ÂüÁ¶Çϼ¼¿ä.

  • Â÷¼¼´ë Capturem¢â membrane ±â¼úÀ» Àû¿ëÇÑ His-tagged protein Á¤Á¦
  • No-waiting workflow: ½Ç¿Â¿¡¼­ 15ºÐ - 30ºÐÀ̸é Á¤Á¦ ¿Ï·á
  • Bed-volumeÀÌ ÀÛ¾Æ °í¼øµµ, °í¼öÀ², °í³óµµÀÇ ´Ü¹éÁú ÃßÃâ °¡´É
  • ºñº¯¼º ¹× º¯¼º Á¶°Ç (EDTA, ȯ¿øÁ¦, ±Û¸®¼¼·Ñ µî)¿¡¼­µµ Á¤Á¦ °¡´É
  • Lysis compatibility: xTractor Buffer, ¶Ç´Â Ÿ»çÀÇ lysis buffer¿¡µµ ȣȯ
  • ¿ë·®º°·Î 4°¡Áö ŸÀÔ Á¦°ø: Miniprep, Maxiprep, 24well plate, 96well plate, Large volume
Á¦Ç°¼³¸í
Capturem¢â His-tagged Purifcation ½Ã¸®Áî´Â Â÷¼¼´ë ¸âºê·¹ÀÎ ±â¼úÀ» Àû¿ëÇÏ¿© His-tagged proteinÀ» Á¤Á¦ÇÒ ¼ö ÀÖ´Â Çõ½ÅÀûÀÎ Á¦Ç°À¸·Î½á, High-capacity Ni-NTA Capturem¢â membranesÀÌ Â÷¿ëµÇ¾îÀÖ´Ù. ±âÁ¸ÀÇ metal affinity resin¿¡ ºñÇÏ¿© º¸´Ù ´Ù¾çÇÑ »ùÇà (¹ÚÅ׸®¾Æ, Æ÷À¯·ù Àû¿ë °¡´É)°ú ´Ù¾çÇÑ Á¶°Ç (ºñº¯¼º ¶Ç´Â º¯¼º Á¶°Ç (8M urea, 6M Guanidine-HCl)¿¡¼­µµ Àû¿ëÀÌ °¡´ÉÇÏ´Ù.
Capturem¢â ½Ã¸®Áî´Â ·¹ÁøÀ» ÀÌ¿ëÇÑ Á¤Á¦¿Í´Â ´Þ¸® ºü¸¥ ½Ã°£ ³»¿¡ ½Ç¿Â¿¡¼­ Á¶ÀÛÇϱ⠶§¹®¿¡ ´Ü¹éÁúÀÇ ºÐÇØ(degradation), ÀÀÁý (aggregation), ½ÇȰ (loss of activity) µîÀÇ ´Ü¹éÁú
¿­º¯¼º, ºÒȰ¼ºÈ­¿¡ ´ëÇÑ À§ÇèÀ» °¨¼Ò½Ãų ¼ö ÀÖ´Ù. ¶ÇÇÑ Bed volumeÀÌ ÀÛ°í (2ul: Miniprep), Binding capacity°¡ ³ô¾Æ ( 75 mg/cm3), °í¼øµµ, °í³óµµÀÇ ´Ü¹éÁúÀ» Á¤Á¦ÇÒ ¼ö ÀÖ´Ù. Capturem¢â His-tagged Purifcation ½Ã¸®Áî´Â ¾Æ·¡¿Í °°ÀÌ Miniprep, Maxiprep, 24well plate, 96well plate, Large volumeÀ¸·Î Á¦°øµÈ´Ù (Ç¥ 1).

Ç¥ 1. Capturem¢â His-tagged protein purification kit ½Ã¸®Áî ºñ±³

 

Miniprep Kit

Maxiprep Kit

96well plate

24well plate

Large volume

Code

635710

635713

635714

635730

635724

ÇüÅÂ

Mini Spin Column
Miniprep Column

Maxiprep Nickel Column
Maxiprep column

96 Nickel Plate
96 Nickel plate


24 Nickel plate

Large Volume Unit
Large volume unit

Input sample

~ 800 §¡

~ 25 §¢

~ 1,000 §¡

~ 4 §¢

150 ~ 500 §¢

¿ë·®

20 ȸ

6 ȸ

96 well

24 well

4 ȸ

¼Ò¿ä½Ã°£

5 ºÐ

15 ºÐ

15 ºÐ

15ºÐ

15 ~ 30 ºÐ

Á¤Á¦³óµµ

0.3 ~ 1 §·/§¢

1.6 ~ 4.5 §·/§¢

0.3 ~ 1 §·/§¢

~ 0.8 §·/1§¢

up to 2.0 §·/§¢

Á¤Á¦¼öÀ²***

80§¶/column

1.5 §·/tube

80§¶/well

800§¶/well

10~25 §·/unit

Centrifuge
Á¶°Ç

11,000 x g, 1ºÐ

2,000 x g, 3ºÐ*

2,000 x g, 3ºÐ*

600 x g, 2ºÐ*

X

Vacuum ¿©ºÎ

X

X

O

O

O**

Buffer
Æ÷ÇÔ ¿©ºÎ

O

O

X

X

X


* Maxiprep Kit¿¡´Â 50§¢ tube¿ë, 24well plate, 96well plate¿¡´Â °¢°¢ 24well plate, 96well plate¿ë centrifuge°¡ ÇÊ¿äÇÕ´Ï´Ù.
** Large volumeÀÇ °æ¿ì, Vacuum source (e.g. vacuum pump equipped with a trap and a vacuum controller, or a peristaltic pump)°¡ ÇÊ¿äÇÏ´Ù.
*** Á¤Á¦¼öÀ²Àº »ç¿ëÇϰíÀÚ ÇϽô Sample type, species, antibody isotype¿¡ µû¶ó ´Þ¶óÁú ¼ö ÀÖ½À´Ï´Ù.


±×¸² 1. Capturem His-Tagged Purification ½Ã¸®ÁîÀÇ Workflow
xTractor Buffer¸¦ Capturem¢â Ä÷³¿¡ ÷°¡ÇÏ¿© EquilibrationÀ» ÁøÇà ÈÄ, His-tagged proteinÀ» ÷°¡ÇÏ°í °áÇÕ½ÃŲ´Ù. µÚÀ̾î Washing Buffer¸¦ ÀÌ¿ëÇÏ¿© ¼¼Ã´ ¹× ¿ø½ÉºÐ¸® ÈÄ
Elution Buffer¸¦ ÀÌ¿ëÇÏ¿© ¿ëÃâÇÑ´Ù. Miniprep°ú 96well plate ŸÀÔÀÇ °æ¿ì´Â Elution buffer 100ul¸¦, MaxiprepÀÇ °æ¿ì´Â Elution buffer 500ul¸¦ »ç¿ëÇÏ¿© 90%ÀÌ»ó Elution °¡´ÉÇÏ´Ù.

Ç¥ 2. Capturem His-Tagged Purification ½Ã¸®ÁîÀÇ BufferÀÇ Á¶¼º
Buffer

Á¶¼º

Lysis Buffer

xTractor Buffer (Code 635625) »ç¿ë ±ÇÀå. Standard lysis bufferµµ »ç¿ë °¡´É.

Wash Buffer

150 mM NaCl , 20 mM Na3PO4, pH 7.6

Elution Buffer

500 mM NaCl , 20 mM Na3PO4, 500 mM imidazole, pH7.6



±×¸² 2. Capturem¢â His-tagged Purification Á¦Ç°ÀÇ Membrane technology
Àå½Ã°£À» ÇÊ¿ä·Î ÇÏ´Â ·¹Áø Á¤Á¦¿¡ ºñÇØ, Capturem¢â membraneÀº Ç¥¸éÀû°ú ¿ë·® (Capacity)°¡ Å©±â ¶§¹®¿¡ ºü¸¥ ½Ã°£ ³»¿¡ ¸· Åë°ú°¡ °¡´ÉÇϹǷΠ´Ü¹éÁú º¯¼ºÀÌ ¾ø´Â ´Ü¹éÁúÀ» Á¤Á¦ °¡´ÉÇÏ´Ù. ¶ÇÇÑ ³·Àº ¾Ð·ÂÀ¸·Î ¿ë¾×ÀÌ Åë°úÇÒ ¼ö ÀÖÀ¸¹Ç·Î ´Ü¹éÁúÀÇ Ç°Áú ÀúÇϸ¦ ¹æÁöÇϸç, Bed volumeÀÌ ÀÛ¾Æ °í¼øµµ·Î Á¤Á¦ÇÒ ¼ö ÀÖ´Ù.

Ç¥ 3. Reagents Compatible with His-Tagged Purification Miniprep Kits
Reagent

Compatibility

MOPS

200 mM

HEPES

200 mM

Tris

200 mM

EDTA

10 mM

Beta-mercaptoethanol

30 mM

DTT

10 mM

TCEP

5 mM

Guanidine-HCl

6 M

Urea

8 M

Nonionic detergent (Triton X-100)

2%

Nonionic detergent (Tween 20)

2%

Anionic detergent (SDS)

1%

Arginine

500 mM

Glycine

100 mM

Histidine

20 mM

Sodium chloride

2 M

Imidazole

40 mM

Glycerol

10%



±×¸² 3. Æ÷À¯·ù¼¼Æ÷¿¡¼­ ¹ßÇö½ÃŲ his-tagged protein Á¤Á¦
293T cell¿¡ 6x His-tagged MAPK1 ¶Ç´Â ADRB2 À¯ÀüÀÚ¸¦ Æ÷ÇÔÇÑ Çö󽺹̵带 Transfection ÈÄ, Capturem¢â His-Tagged Purification Miniprep KitÀ» ÀÌ¿ëÇÏ¿© ´Ü¹éÁúÀ» Á¤Á¦ÇÏ¿´´Ù. EquilibrationÀ» ¿Ï·áÇÑ Ä÷³¿¡ Lysate ó¸®, Washing ÈÄ Elution Buffer 300ul·Î ´Ü¹éÁúÀ» ¿ëÃâÇϰí 4-20% Polyacrylamide gel Àü±â¿µµ¿ ¹× Western blottingÀ» ÁøÇàÇÏ¿´´Ù.
Ponceau S stainingÀ» ÅëÇÏ¿© membrane ³»ÀÇ ¸ðµç ´Ü¹éÁúÀÌ °ËÃâµÇ´Â °ÍÀ» È®ÀÎÇÒ ¼ö ÀÖ¾úÀ¸¸ç, MAPK, ADRB2 ƯÀÌÀûÀÎ Ç×ü¸¦ »ç¿ëÇÑ Western blot¿¡¼­ °¢°¢ÀÇ Á¤Á¦µÈ ´Ü¹éÁúÀ»
È®ÀÎÇÒ ¼ö ÀÖ¾ú´Ù.


±×¸² 4. Capturem¢â His-tagged Purification Large Volume
(¿ÞÂÊ) Capturem His-Tagged Purification Large Volume Units (¿À¸¥ÂÊ) Bottle Thread Adaptor


±×¸² 5. Capturem¢â His-Tagged Purification Large Volume Unit Àû¿ë»çÁø
(A) 33mm Áö¸§ÀÇ ÀÔ±¸¸¦ °¡Áø bottle¿¡ ¼³Ä¡ÇÑ Capturem His-Tagged Purification Large Volume Unit
(B) 6x His-GFPuv ´Ü¹éÁúÀ» Æ÷ÇÔÇÑ cell lysate¸¦ Capturem unit¿¡ ·ÎµùÇÑ »çÁø
(C, À§) 6x His-GFPuv ´Ü¹éÁúÀ» Æ÷ÇÔÇÑ cell lysate ·Îµù ÈÄ, washingÀ» ÇÑ membraneÀÇ ¸ð½À. Ȳ·Ï»öÀ» ¶ç°í ÀÖ´Ù.
(C, ¾Æ·¡) Elution ÁøÇà ÈÄÀÇ membraneÀÇ »ö. ¿ø·¡ÀÇ membrane »öÀÎ Èò»öÀ» ¶ç°í ÀÖ´Ù.
(D) 50§¢ tube¿¡ elutionÀ» ¿Ï·áÇÑ 6x His-GFPuv ´Ü¹éÁú ¿ë¾×
Àû¿ë
  • Biochemical/enzymatic assays
  • Pull-down or other protein interaction assays
  • Confirmation screening
  • Protein/ligand affinity analysis
  • Crystallography or other structural analyses
  • Antibody production
  • Toxicology studies
  • Protein transfer to live cells
±¸¼ºÇ° (ÀÚ¼¼ÇÑ ³»¿ëÀº CoA¸¦ ÂüÁ¶Çϼ¼¿ä)
Capturem His-Tagged Purification Miniprep Kit
Mini Spin Column

20°³

xTractor Buffer

15ml¡¿ 2

Wash Buffer

10 ml

Elution Buffer

10 ml


Capturem His-Tagged Purification Maxiprep Kit
Capturem Maxiprep Nickel Column

6°³

Capturem Maxiprep Filters

6°³

xTractor Buffer

200 ml

Wash Buffer

40 ml

Elution Buffer

15 ml


Capturem His-Tagged Purification Maxiprep Columns
Capturem Maxiprep Nickel Column 25°³¡¿2

Capturem His-Tagged Purification 96
Capturem 96 Nickel Plate 1Àå

Capturem¢â His-Tagged Purification 24-Well Plate
Capturem His-Tagged Purification 24-Well Plate 1Àå

Capturem His-Tagged Purification Large Volume
Capturem His-Tagged Purification Large Volume Unit 4°³(2 Units per box)
Bottle Thread Adaptor 1°³

£ªCapturem His-Tagged Purification Maxiprep Columns, Capturem His-Tagged Purification Large Volume ¹× Capturem His-Tagged Purification 96, Capturem¢â His-Tagged Purification 24-Well Plate¿¡´Â Buffer°¡ Æ÷ÇԵǾîÀÖÁö
¾Ê½À´Ï´Ù.
ÁÖ¿ä »ùÇÿ¡ µû¸¥ xTractor Buffer Lysis Protocol

Àû¿ë»ùÇà Á¾

Protocol (Ŭ¸¯ÇÏ¿© ´Ù¿î·ÎµåÇϼ¼¿ä)

xTractor Buffer Ç¥ÁØ ¸Å´º¾ó

xTractor Buffer and xTractor Buffer Kit User Manual 

Bacteria

xTractor Buffer Bacterial Sample Preparation Protocol-At-A-Glance 

Baculovirus

xTractor Buffer Baculovirus Sample Preparation Protocol-At-A-Glance 

Mammalian

xTractor Buffer Mammalian Sample Preparation Protocol-At-A-Glance 

Yeast

xTractor Buffer Yeast Sample Preparation Protocol-At-A-Glance 







Keyword : His-tagged protein,È÷½ºÅÃ,His-tag protein,His-tag,´Ü¹éÁú Á¤Á¦,´Ü¹éÁú,Capturem,Á¤Á¦
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