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ApopTag¢ç Peroxidase In Situ Apoptosis Detection Kit

Apoptosis¸¦ ¾î¶² ¹æ¹ýÀ¸·Î ÃøÁ¤ÇÏ´Â °Ç°¡¿ä?
TUNEL assay ¹æ¹ýÀ» ÀÌ¿ëÇÏ¿© DNA frgment ¸»´Ü¿¡ digoxigenin-dNTP¸¦ ºÙÈù ÈÄ, Anti-digoxigenin-HRP°ú ¹ÝÀÀ½ÃÄÑ DABÀ¸·Î ¹ß»öÇÑ µÚ, Çö¹Ì°æÀ¸·Î apoptosis¸¦ °üÂûÇÕ´Ï´Ù.
Cell¿¡¼­ ¹Ù·Î »ç¿ëÇÒ ¼ö ÀÖ³ª¿ä?
³×. Cell cultures »ùÇà ¿Ü¿¡µµ formalin-fixed tissues, paraffin embedded tissues, cryostat sections, cell suspensions, cytospins µî cell¸¸ ÀÖÀ¸¸é ¹Ù·Î apoptosis È®ÀÎÀÌ ½±°Ô °¡´ÉÇÕ´Ï´Ù.
Áٱ⼼Æ÷ÀÇ apoptosis È®Àεµ °¡´ÉÇѰ¡¿ä?
³×, °¡´ÉÇÕ´Ï´Ù. º¸´Ù ÀÚ¼¼ÇÑ ³»¿ëÀº ´ÙÀ½ÀÇ ÆÄÀÏ(an1237en00.pdf)À» Âü°íÇØÁÖ¼¼¿ä.
XyleneÀº ¾ðÁ¦ »ç¿ëÇϳª¿ä?
Ç¥º» ¼³Ä¡ Àü de-waxing paraffin tissues¿Í ºÐ¼® ¸¶Áö¸· ´Ü°èÀÎ dehydration °úÁ¤¿¡¼­ XyleneÀÌ »ç¿ëµË´Ï´Ù. XyleneÀº À¯±â¿ë¸Å·Î »ç¿ëÇÏÁö ¾ÊÀ» ¶§´Â ¶Ñ²±À» ²À ´ÝÀ¸¼Å¾ß ÇÕ´Ï´Ù.
Control Slidesµµ ÇÔ²² Á¦°øµÇ³ª¿ä?
º» Kit ±¸¼ºÇ° Áß Control Slides´Â Æ÷ÇÔµÇÁö ¾Ê½À´Ï´Ù. ApopTag¢ç Positive Control Slides (Cat. No. S7115)¸¦ º°µµ·Î ±¸¸ÅÇØÁÖ¼¼¿ä.
ApopTag¢ç Plus Peroxidase In Situ Apoptosis Detection Kit (S7101)°ú ApopTag¢ç Plus Fluorescein In Situ Apoptosis Detection Kit(S7111)´Â Positive control slides°¡ ÇÔ²² Á¦°øµË´Ï´Ù.
ºÐ¼® ÁøÇà µµÁß ½ÇÇèÀ» Àá½Ã ¸ØÃß°í ½Í½À´Ï´Ù. ¾î´À °úÁ¤¿¡¼­ Áß´ÜÇÏ¿© »ùÇÃÀ» Àӽ÷Πº¸°üÇÒ ¼ö ÀÖÀ»±î¿ä?
  1. ½½¶óÀ̵带 Equilibration Buffer ¶Ç´Â water·Î Àû½Å ÈÄ, 4¡ÆC-»ó¿Â¿¡¼­ ¾à 60ºÐ°£ º¸°üÇÒ ¼ö ÀÖ½À´Ï´Ù.
  2. TdT enzyme°ú ¹ÝÀÀ½ÃŲ ÀÌÈÄ : ½½¶óÀ̵带 Stop-Wash SolutionÀ¸·Î 5ºÐ°£ washÇϰí, 70% EtOH°¡ µé¾îÀÖ´Â coplin jar¿¡ ³Ö¾î -20¡ÆC¿¡¼­ ÃÖ¼Ò 3ÀÏ º¸°üÀÌ °¡´ÉÇÕ´Ï´Ù.
    º¸°ü ÈÄ ºÐ¼®À» °è¼Ó ÁøÇàÇϱâ Àü, PBS·Î 2ºÐ°£°ÝÀ¸·Î »ùÇÃÀ» 3¹ø washingÇÏ¿© À̹°ÁúÀ» Á¦°ÅÇØÁÖ¼¼¿ä.
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»ùÇÿ¡ µû¶ó ºÐ¼®½Ã°£Àº Á¶±Ý Â÷À̰¡ ÀÖ½À´Ï´Ù. Paraffin sectionsÀÇ °æ¿ì 4½Ã°£, Tissue cryosections ¶Ç´Â cultured cellsÀÇ °æ¿ì ¾à 3½Ã°£ ¹Ý Á¤µµÀÇ ½Ã°£ÀÌ °É¸³´Ï´Ù.
ºÐ¼®½Ã°£À» ÁÙÀ̱â À§Çؼ± Pretreatment¿Í quenching ÈÄ, »ùÇÃÀ» deionized water(dH2O)·Î washing ÇØÁÖ¼¼¿ä. TdT ¶óº§¸µ Àü, Equilibration Buffer·Î washing ÇÏ´Â °úÁ¤À» ÁÙÀÏ ¼ö ÀÖ½À´Ï´Ù.