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Retrovirus ³óÃà

-

Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
ºñ°í »ç¿ëÀڸŴº¾ó
Clontech
631455
Retro-X Concentrator
°ü·ÃÇмú ±¸¸ÅÇϱâ
100 ml
494,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x33007
Clontech
631456
Retro-X Concentrator
°ü·ÃÇмú ±¸¸ÅÇϱâ
500 ml
1,706,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â

  • ÃÊ°í¼Ó¿ø½ÉºÐ¸®±â ¾øÀÌ ¹ÙÀÌ·¯½º ¿ª°¡¸¦ 100 ¹è±îÁö Áõ°¡
  • °£´ÜÇÑ Á¶ÀÛ °úÁ¤ : mix, wait ±×¸®°í spin
  • ¹ø°Å·Î¿ò ¾øÀÌ ºÎÇǸ¦ ½±°Ô scale up °¡´É
Retro-X Concentrator·Î retrovirusÀÇ ¿ª°¡¸¦ 100 ¹è±îÁö Áõ°¡½Ãų ¼ö ÀÖÀ¸¸ç, ´õ ¸¹Àº ¹ÙÀÌ·¯½º¸¦ ¸¸µé°Å³ª, Ãß°¡·Î packaging ¼¼Æ÷¿¡ transfectionÇÒ ÇÊ¿ä ¾øÀÌ ÃæºÐÈ÷ ³ôÀº MOI·Î
¸ñÀû¼¼Æ÷¸¦ °¨¿°½Ãų ¼ö ÀÖ´Ù.
Simple Á¶ÀÛ°úÁ¤: Mix, Wait ±×¸®°í Spin
Retro-X Concentrator´Â ºü¸£°í, °£ÆíÇÏ¸ç ¾î¶°ÇÑ Á¾·ùÀÇ retrovirus stockµµ ³óÃà½Ãų ¼ö ÀÖ´Â ¾ÆÁÖ È¿À²ÀûÀÎ ¹æ¹ýÀ» Á¦°øÇÑ´Ù. ¿ì¼± retrovirus »óÃþ¾×À» Retro-X Concentrator ½Ã¾à°ú ¼¯Àº ÈÄ ÇÏ·í¹ã ¹ÝÀÀ½ÃŲ ÈÄ ¹ÝÀÀ¾×À» ÀÏ¹Ý ¿ø½ÉºÐ¸®±â·Î ¿ø½ÉºÐ¸®ÇÑ´Ù (Figure 1). »óÃþ¾×ÀÇ ¾ç°ú viral envelope¿¡ µû¶ó MMLV ¶Ç´Â MSCV¿¡ ±â¹ÝÀ» µÐ retroivirusÀÇ ¿ª°¡¸¦
100 ¹è±îÁö Áõ°¡½Ãų ¼ö ÀÖÀ¸¸ç (Figure 2), ³ôÀº ȸ¼öÀ²À» ¾òÀ» ¼ö ÀÖ´Ù. ClontechÀÇ Retro-XSystemÀÇ ¸ðµç retroiviral vector »Ó¸¸ ¾Æ´Ï¶ó ´Ù¸¥ ¸ðµç retroivirusÀÇ »óÃþ¾×À»
Retro-X Concentrator·Î ³óÃàÇÒ ¼ö ÀÖÀ¸¸ç ÇÊ¿ä¿¡ µû¶ó ¿ë·®À» Áõ°¨ÇÒ ¼ö ÀÖ´Ù.
Table I: Retro-X Concentrator vs. Ultracentrifugation
Feature Retro-X Concentrator Ultracentrifugation
Easily Scalable Yes No
Specialized Equipment No Yes
Ease-of-Use ++++ +
100 ¹è±îÁö ¿ª°¡ Áõ°¡
Retro-X ConcentratorÀ» »ç¿ëÇÏ¸é ´Ù¾çÇÑ envelope-pseudotyped retroivirusÀÇ »óÃþ¾× 10 mlÀ» 100 ¥ìI·Î ³óÃàÇÏ¿© ¿ª°¡¸¦ 46~116 ¹è±îÁö Áõ°¡½Ãų ¼ö ÀÖ´Ù (Figure 2).
ÃÊ°í¼Ó¿ø½ÉºÐ¸®º¸´Ù ´õ¿í °£´ÜÇÑ ¹æ¹ý
Retro-X Concentrator°ú ÃÊ°í¼Ó¿ø½ÉºÐ¸® ¹æ¹ýÀ» ºñ±³ÇÏ¿´À» °æ¿ì, Retro -X ¹æ¹ýÀÇ ÀåÁ¡Àº ¸Å¿ì ¸íÈ®ÇÏ´Ù (Table 1). Retro-X Concentrator´Â ÃÊ°í¼Ó¿ø½ÉºÐ¸® ¹æ¹ýº¸´Ù ´õ ½±°í À¶Å뼺ÀÌ ÀÖ´Ù. Retro-X Concentrator ½Ã¾àÀº 4x ³óµµÀÇ ½Ã¾àÀ¸·Î Á¦°øµÇ±â ¶§¹®¿¡ »óÃþ¾×¿¡ Æ÷ÇÔµÈ ¹ÙÀÌ·¯½ºÀÇ ¾ç°ú ¿ª°¡¿¡ »ó°ü¾øÀÌ »ç¿ëÇÒ ¼ö ÀÖ´Ù. »óÃþ¾× 250 ml ±îÁöµµ Àû¿ë °¡´ÉÇÏ´Ù.


Figure 1. The Retro-X Concentrator protocol. Add Retro-X Concentrator reagent to clarified viral supernatant, incubate overnight at 4¡ÆC, and spin. That¡¯s it.


Figure 2. Retro-X Concentrator increases virus titer by up to 100-fold. Retrovirus expressing DsRed2 was packaged using the Retro-X Universal Packaging System and four different viral envelopes. Retro-X Concentrator was used to concentrate dualtropic (10A1), amphotropic (Ampho), ecotropic (Eco), and VSV-G (pantropic) pseudotyped retrovirus samples from 10 ml down to 100 ¥ìl (Panels A & B). Crude and concentrated viral stocks were then titrated on HT1080 or NIH 3T3 cells and analyzed via flow cytometry at 72 hr postinfection to determine the percentage of transduced cells.

Components & Storage Conditions
°¢ Á¦Ç° ±¸¼º¹°°ú º¸°üÁ¶°ÇÀº Certificate of Analysis ¸¦ ÂüÁ¶ÇϽʽÿÀ.

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