[原创]免疫共沉淀反应 （Protein A Agarose）试剂盒
<p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><font face="Times New Roman"><b style="mso-bidi-font-weight: normal;"><span lang="EN-US" style="FONT-SIZE: 16pt;">Immunoprecipitation Kit<span style="mso-spacerun: yes;"> </span></span></b><span lang="EN-US"><font size="3">Protein A Agarose</font></span></font></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><font size="3"><span style="FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman";">免疫共沉淀反应</span><font face="Times New Roman"></font><span style="FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman";">（</span><span lang="EN-US"><font face="Times New Roman">Protein A Agarose</font></span><span style="FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman";">）试剂盒</span></font></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><b style="mso-bidi-font-weight: normal;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana;">Description: <p></p></span></b></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><b style="mso-bidi-font-weight: normal;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana;"><p> </p></span></b></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana;">Anti-FLAG affinity gel is a purified mouse IgG<sub>1</sub> monoclonal antibody covalently conjugated to agarose by hydrazide linkage. The kit is designed for purification or immunoprecipitation of FLAG-tagged proteins from a variety of sources including mammalian cells, insect cells, bacteria and plant.<p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana;"><p> </p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><b style="mso-bidi-font-weight: normal;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana;">Size: </span></b><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana;">30 standard assays<p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana;"><p> </p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><b style="mso-bidi-font-weight: normal;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana;">Kit components:<p></p></span></b></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana;"><p> </p></span></p><table class="MsoTableGrid" cellspacing="0" cellpadding="0" border="0" style="MARGIN: auto auto auto 27pt; BORDER-COLLAPSE: collapse; mso-padding-alt: 0cm 5.4pt 0cm 5.4pt; mso-yfti-tbllook: 480;"><tbody><tr style="mso-yfti-irow: 0; mso-yfti-firstrow: yes;"><td valign="top" width="113" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: windowtext 1pt solid; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 84.5pt; PADDING-TOP: 0cm; BORDER-BOTTOM: windowtext 1pt solid; BACKGROUND-COLOR: transparent; mso-border-bottom-alt: solid windowtext .5pt; mso-border-top-alt: solid windowtext .5pt;"><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana;">Components<p></p></span></p></td><td valign="top" width="202" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: windowtext 1pt solid; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 151.8pt; PADDING-TOP: 0cm; BORDER-BOTTOM: windowtext 1pt solid; BACKGROUND-COLOR: transparent; mso-border-bottom-alt: solid windowtext .5pt; mso-border-top-alt: solid windowtext .5pt;"><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana;">Name<p></p></span></p></td><td valign="top" width="83" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: windowtext 1pt solid; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 62.5pt; PADDING-TOP: 0cm; BORDER-BOTTOM: windowtext 1pt solid; BACKGROUND-COLOR: transparent; mso-border-bottom-alt: solid windowtext .5pt; mso-border-top-alt: solid windowtext .5pt;"><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana;">Cat#<p></p></span></p></td><td valign="top" width="80" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: windowtext 1pt solid; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 60.35pt; PADDING-TOP: 0cm; BORDER-BOTTOM: windowtext 1pt solid; BACKGROUND-COLOR: transparent; mso-border-bottom-alt: solid windowtext .5pt; mso-border-top-alt: solid windowtext .5pt;"><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana;">Size<p></p></span></p></td></tr><tr style="mso-yfti-irow: 1;"><td valign="top" width="113" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 84.5pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent; mso-border-top-alt: solid windowtext .5pt;"><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana;">Component A<p></p></span></p></td><td valign="top" width="202" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 151.8pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent; mso-border-top-alt: solid windowtext .5pt;"><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US"><font face="Times New Roman">Protein A Agarose</font></span></p></td><td valign="top" width="83" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 62.5pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent; mso-border-top-alt: solid windowtext .5pt;"><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US"><font face="Times New Roman">IR003</font></span></p></td><td valign="top" width="80" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 60.35pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent; mso-border-top-alt: solid windowtext .5pt;"><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US"><font face="Times New Roman">1mL</font></span></p></td></tr><tr style="mso-yfti-irow: 2;"><td valign="top" width="113" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 84.5pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;"><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana;">Component B<p></p></span></p></td><td valign="top" width="202" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 151.8pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;"><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US"><font face="Times New Roman">Binding Buffer</font></span></p></td><td valign="top" width="83" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 62.5pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;"><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US"><font face="Times New Roman">MP011</font></span></p></td><td valign="top" width="80" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 60.35pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;"><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US"><font face="Times New Roman">50mL</font></span></p></td></tr><tr style="mso-yfti-irow: 3;"><td valign="top" width="113" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 84.5pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;"><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana;">Component C<p></p></span></p></td><td valign="top" width="202" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 151.8pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;"><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US"><font face="Times New Roman">Washing Buffer (5X)</font></span></p></td><td valign="top" width="83" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 62.5pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;"><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US"><font face="Times New Roman">MP011T</font></span></p></td><td valign="top" width="80" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 60.35pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;"><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US"><font face="Times New Roman">50mL</font></span></p></td></tr><tr style="mso-yfti-irow: 4;"><td valign="top" width="113" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 84.5pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;"><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana;">Component D<p></p></span></p></td><td valign="top" width="202" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 151.8pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;"><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US"><font face="Times New Roman">Elution Buffer</font></span></p></td><td valign="top" width="83" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 62.5pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;"><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US"><font face="Times New Roman">MP005</font></span></p></td><td valign="top" width="80" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 60.35pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;"><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US"><font face="Times New Roman">1.5mL</font></span></p></td></tr><tr style="mso-yfti-irow: 5; mso-yfti-lastrow: yes;"><td valign="top" width="113" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 84.5pt; PADDING-TOP: 0cm; BORDER-BOTTOM: windowtext 1pt solid; BACKGROUND-COLOR: transparent; mso-border-bottom-alt: solid windowtext .5pt;"><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana;">Component E<p></p></span></p></td><td valign="top" width="202" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 151.8pt; PADDING-TOP: 0cm; BORDER-BOTTOM: windowtext 1pt solid; BACKGROUND-COLOR: transparent; mso-border-bottom-alt: solid windowtext .5pt;"><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US"><font face="Times New Roman">Neutralization Buffer</font></span></p></td><td valign="top" width="83" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 62.5pt; PADDING-TOP: 0cm; BORDER-BOTTOM: windowtext 1pt solid; BACKGROUND-COLOR: transparent; mso-border-bottom-alt: solid windowtext .5pt;"><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US"><font face="Times New Roman">MC030.2</font></span></p></td><td valign="top" width="80" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 60.35pt; PADDING-TOP: 0cm; BORDER-BOTTOM: windowtext 1pt solid; BACKGROUND-COLOR: transparent; mso-border-bottom-alt: solid windowtext .5pt;"><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US"><font face="Times New Roman">0.5mL</font></span></p></td></tr></tbody></table><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana;"><p> </p></span></p> <p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><b style="mso-bidi-font-weight: normal;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana;">Precedures:<p></p></span></b></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><b style="mso-bidi-font-weight: normal;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana;"><p> </p></span></b></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; mso-list: l2 level2 lfo1; tab-stops: list 18.0pt;"><b style="mso-bidi-font-weight: normal;"><span lang="EN-US" style="mso-bidi-font-size: 10.0pt; mso-fareast-font-family: "Times New Roman";"><span style="mso-list: Ignore;"><font face="Times New Roman">A.<span style="FONT: 7pt "Times New Roman";"> </span></font></span></span></b><b style="mso-bidi-font-weight: normal;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana;">Preparation of Cell Lysates (for adherent </span></b><b style="mso-bidi-font-weight: normal;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;">mammalian cells)<p></p></span></b></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;"><p> </p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; mso-list: l1 level1 lfo2; tab-stops: list 18.0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">1.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;">Remove the growth medium from the cells to be analyzed. Rinse the cells twice with PBS buffer (Cat# CC008).<p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; mso-list: l1 level1 lfo2; tab-stops: list 18.0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">2.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;">Add 10ml (10-cm plate), scrape the cells off the plate and transfer cells into 15-cm Folcon tube.<p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; mso-list: l1 level1 lfo2; tab-stops: list 18.0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">3.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;">Centrifuge the sample at 1000 x g for 5 mins.<p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; mso-list: l1 level1 lfo2; tab-stops: list 18.0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">4.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;">Discard the PBS, add lysis buffer (Cat# MP011T) (10<sup>6</sup>-10<sup>7</sup> cells/ml).<p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; mso-list: l1 level1 lfo2; tab-stops: list 18.0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">5.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;">Incubate the cells for 15-30 minutes on a shaker.<p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; mso-list: l1 level1 lfo2; tab-stops: list 18.0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">6.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;">Centrifuge the cell lysate for 10 minutes at 12,000 x g.<p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; mso-list: l1 level1 lfo2; tab-stops: list 18.0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">7.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;">Transfer the supernatant to a 1.5ml eppendorf tube.<p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; mso-list: l1 level1 lfo2; tab-stops: list 18.0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">8.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;">For immediate use, keep on ice. If the supernatant is not to be used immediately, store it at –<chmetcnv wst="on" unitname="ﾰC" sourcevalue="70" hasspace="True" negative="False" numbertype="1" tcsc="0">70 °C</chmetcnv>.<p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;"><p> </p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; mso-list: l2 level2 lfo1; tab-stops: list 18.0pt;"><b style="mso-bidi-font-weight: normal;"><span lang="EN-US" style="mso-bidi-font-size: 10.0pt; mso-fareast-font-family: "Times New Roman";"><span style="mso-list: Ignore;"><font face="Times New Roman">B.<span style="FONT: 7pt "Times New Roman";"> </span></font></span></span></b><b style="mso-bidi-font-weight: normal;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;">Immunoprecipitation of FLAG-tagged Proteins<p></p></span></b></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;"><p> </p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; mso-list: l0 level1 lfo3; tab-stops: list 18.0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">1.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;">Thoroughly suspend the Anti-FLAG affinity agarose beads.<p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; mso-list: l0 level1 lfo3; tab-stops: list 18.0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">2.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;">Transfer 30ul of the gel suspension to a 1.5ml eppendorf tube. (For beads transfer, use plastic pipette tip with the end cut for about <chmetcnv wst="on" unitname="mm" sourcevalue="2" hasspace="False" negative="False" numbertype="1" tcsc="0">2mm</chmetcnv> to allow the beads to be transferred).<p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; mso-list: l0 level1 lfo3; tab-stops: list 18.0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">3.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;">Centrifuge the beads briefly to bring the beads to the bottom of the tube.<p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; mso-list: l0 level1 lfo3; tab-stops: list 18.0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">4.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;">Wash the beads twice with 0.5 ml 1X Washing Buffer.<p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; mso-list: l0 level1 lfo3; tab-stops: list 18.0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">5.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;">Add 500-1000ul of cell lysates (up to 1mg) to the beads. The lysates could be diluted with Binding Buffer.<p></p></span></p> <p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; mso-list: l2 level2 lfo1; tab-stops: list 18.0pt;"><b style="mso-bidi-font-weight: normal;"><span lang="EN-US" style="mso-bidi-font-size: 10.0pt; mso-fareast-font-family: "Times New Roman";"><span style="mso-list: Ignore;"><font face="Times New Roman">A.<span style="FONT: 7pt "Times New Roman";"> </span></font></span></span></b><b style="mso-bidi-font-weight: normal;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;">Elution<p></p></span></b></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;"><p> </p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><i style="mso-bidi-font-style: normal;"><u><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;">Elution with <chmetcnv wst="on" unitname="m" sourcevalue=".1" hasspace="True" negative="False" numbertype="1" tcsc="0">0.1 M</chmetcnv> glycine HCl, pH 3.5<p></p></span></u></i></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><i style="mso-bidi-font-style: normal;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;"><p> </p></span></i></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; mso-list: l1 level1 lfo2; tab-stops: list 18.0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">1.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;">Add 100ul Elution Buffer to each sample.<p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; mso-list: l1 level1 lfo2; tab-stops: list 18.0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">2.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;">Incubate the samples and controls with gentle shaking for 5 minutes at room temperature.<p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; mso-list: l1 level1 lfo2; tab-stops: list 18.0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">3.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;">Centrifuge the beads for 30 seconds at 5,000 x g. Transfer the supernatants to a new tube containing 10ul Neutralization Buffer.<p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><i style="mso-bidi-font-style: normal;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;">Note: The procedure should be performed at room temperature. Do not leave the beads in this buffer >20 minutes.<p></p></span></i></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;"><p> </p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><i style="mso-bidi-font-style: normal;"><u><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;">Elution with SDS-PAGE Sample Loading Buffer<p></p></span></u></i></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;"><p> </p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; mso-list: l0 level1 lfo3; tab-stops: list 18.0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">1.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;">Add 20ul of 2X sample loading buffer (Cat# MP006.1) to each sample.<p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; mso-list: l0 level1 lfo3; tab-stops: list 18.0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">2.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;">Boil the samples for 5 minutes.<p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; mso-list: l0 level1 lfo3; tab-stops: list 18.0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">3.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;">Briefly votex the tube and centrifuge the samples at 5,000 x g for 30 seconds to pellet agarose. <p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; mso-list: l0 level1 lfo3; tab-stops: list 18.0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">4.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;">Transfer the supernatants to a new tube. <p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; mso-list: l0 level1 lfo3; tab-stops: list 18.0pt;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-fareast-font-family: Verdana; mso-bidi-font-family: Verdana;"><span style="mso-list: Ignore;">5.<span style="FONT: 7pt "Times New Roman";"> </span></span></span><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;">The samples are ready for loading on SDS-PAGE and immunoblotting using Anti-FLAG or specific antibodies against the fusion protein or associated proteins. <p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><i style="mso-bidi-font-style: normal;"><span lang="EN-US" style="FONT-SIZE: 10pt; FONT-FAMILY: Verdana; mso-bidi-font-family: Arial;">Note: The procedure should be preformed at room temperature. Sample buffer should be at room temperature before use</span></i><b style="mso-bidi-font-weight: normal;"><span lang="EN-US" style="FONT-SIZE: 16pt;"><p></p></span></b></p> <p><span style="FONT-SIZE: 12pt; FONT-FAMILY: 宋体;">迈晨科技（北京）有限公司竭诚为您服务 ！<span lang="EN-US"><p></p></span></span></p><p><span style="FONT-SIZE: 12pt; FONT-FAMILY: 宋体;">详情请登录网站：<span lang="EN-US">www.macgene.com</span>，联系电话：<span lang="EN-US">010-82057786<p></p></span></span></p> 迈晨科技（北京）有限公司竭诚为您服务 ！
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