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 Á¦Ç°¾È³»  Àü±â¿µµ¿   ´Ü¹éÁú Àü±â¿µµ¿  Antibody Erasing Buffer

Phospho Antibody Stripping Solution

membrane blot¿¡¼­ Ç×ü¸¦ ¶¼¾î³»´Âµ¥ »ç¿ëÇÏ´Â ¿ë¾×À¸·Î, À̸¦ ÀÌ¿ëÇÏ¿© µ¿ÀÏÇÑ blot¿¡ ´ëÇØ ´Ù¾çÇÑ Ç×ü·Î ½ºÅ͵ð ÇÒ¼ö ÀÖ½À´Ï´Ù.

chemiluminescence, radioactive iodine, ȤÀº isotopesµîÀ» ÀÌ¿ëÇÑ western blot¿¡ »ç¿ëÇÒ¼ö ÀÖ½À´Ï´Ù.

Cell Biolabs
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  • Western BlotÈÄ blot¿¡¼­ anti-phosphoprotein antibodies¸¦ ¶¼¾î³»´Âµ¥ »ç¿ëÇÕ´Ï´Ù.
  • µ¿ÀÏÇÑ blotÀ» »ç¿ëÇÔÀ¸·Î½á ºñ¿ëÀÌ Àý°¨µÇ¸ç ´Ù¾çÇÑ Ç×ü¿¡ ´ëÇÑ ¹ÝÀÀÀ» °ËÅäÇÒ¼ö ÀÖ½À´Ï´Ù.
  • anti-phosphoprotein antibodies stripping¿¡ ÃÖÀûÈ­ µÇ¾î ÀÖ½À´Ï´Ù.
  • membrane»ó¿¡ ºÎÂøµÈ Ç׿øÀÇ ¼Õ»óÀ» ÁÖÁö¾Ê°í, multiple strippingÀÌ °¡´ÉÇÕ´Ï´Ù.
  • Antibody strippingÀº heating¾øÀÌ ½Ç¿Â¿¡¼­ °¡´ÉÇÕ´Ï´Ù.
  • 75X ³óÃàÀ¸·Î 10ml Á¦°øµË´Ï´Ù.
»ç¿ë¹ý
  • Freshly prepare enough 1X Phospho Antibody Stripping Solution in a vented chemical hood by diluting the 75X stock in dH2O, mix well.
  • Submerge the blot in the 1X Phospho Antibody Stripping Solution and incubate with gentle mixing for 90 to 120 minutes at room temperature. Ensure sufficient volume is used to completely cover the blot.
  • Wash twice with TBST.
  • Test for complete removal of the phospho-antibody by blotting with secondary antibody, followed by ECL detection.
  • After determining that the membrane is properly stripped, the blot is now ready for re-probing with same or different antibodies.
»ç¿ë¿¹
Figure 1. Multiple Stripping of 4G10 Phosphotyrosine Antibody. 15 μg of serum-stimulated COS-7
cell lysate was subject to 4G10 anti-phosphotyrosine western blot. The same blot was stripped with Phospho Antibody Stripping Solution and re-probed with either HRP-secondary conjugate alone (Ctrl) or the primary 4G10 antibody and the HRP-secondary conjugate (4G10), followed by detection with ECL reagents. Films were exposed for 30 seconds for the 4G10 blot and 30 minutes for the control blot.