[原创]β－半乳糖苷酶检测试剂盒

macgene 发表于 2009-1-5 09:44:00

b-Galactosidase Assay Kit   b－半乳糖苷酶检测试剂盒Catalogue：MPK006   Storage: 4－8 ºCKit Components: <table class="MsoTableGrid" cellspacing="0" cellpadding="0" border="0" style="MARGIN: auto auto auto 17.75pt; 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="132" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 99pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">Kit Components</td><td valign="top" width="222" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 166.8pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">Chemicals</td><td valign="top" width="90" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 67.2pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">Size</td><td valign="top" width="90" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 67.2pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">Cat #</td></tr><tr style="mso-yfti-irow: 1;"><td valign="top" width="132" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 99pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">I</td><td valign="top" width="222" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 166.8pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">Extraction Buffer</td><td valign="top" width="90" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 67.2pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">10 mL</td><td valign="top" width="90" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 67.2pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">MC045</td></tr><tr style="mso-yfti-irow: 2;"><td valign="top" width="132" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 99pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">II</td><td valign="top" width="222" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 166.8pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">Assay Buffer (b-gal)</td><td valign="top" width="90" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 67.2pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">60 mL</td><td valign="top" width="90" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 67.2pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">MC048</td></tr><tr style="mso-yfti-irow: 3;"><td valign="top" width="132" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 99pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">III</td><td valign="top" width="222" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 166.8pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">Substrate Solution (ONPG)</td><td valign="top" width="90" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 67.2pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">10 mL</td><td valign="top" width="90" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 67.2pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">MC049</td></tr><tr style="mso-yfti-irow: 4;"><td valign="top" width="132" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 99pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">IV</td><td valign="top" width="222" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 166.8pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">Stop Solution</td><td valign="top" width="90" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 67.2pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">10 mL</td><td valign="top" width="90" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 67.2pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">MC007.1</td></tr><tr style="mso-yfti-irow: 5; mso-yfti-lastrow: yes;"><td valign="top" width="132" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 99pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">V</td><td valign="top" width="222" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 166.8pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">pCMV-b-gal</td><td valign="top" width="90" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 67.2pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">10 mg</td><td valign="top" width="90" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 67.2pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">MN008</td></tr></tbody></table>

macgene 发表于 2009-1-5 09:45:00

Procedures: Preparation of Cell Lysates <p class="MsoNormal" align="left" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; LINE-HEIGHT: 125%; TEXT-ALIGN: left; mso-pagination: widow-orphan; mso-list: l0 level1 lfo1; tab-stops: list 18.0pt;">Ø       Remove the media from the tissue culture plate wells.<p class="MsoNormal" align="left" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; LINE-HEIGHT: 125%; TEXT-ALIGN: left; mso-pagination: widow-orphan; mso-list: l0 level1 lfo1; tab-stops: list 18.0pt;">Ø       Add 100 μl Extraction Buffer into each well to cover the cells.<p class="MsoNormal" align="left" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; LINE-HEIGHT: 125%; TEXT-ALIGN: left; mso-pagination: widow-orphan; mso-list: l0 level1 lfo1; tab-stops: list 18.0pt;">Ø       Incubate the plate on shaker platform at room temperature for 5-10 minutes.<p class="MsoNormal" align="left" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; LINE-HEIGHT: 125%; TEXT-ALIGN: left; mso-pagination: widow-orphan; mso-list: l0 level1 lfo1; tab-stops: list 18.0pt;">Ø       Place the plates on ice and proceed to the next step. Preparation of Reaction Solution Calculate how much Reaction Solution you need (200ml per well of 96-well plate). Below is the table for preparation of 100 assays. <table class="MsoTableGrid" cellspacing="0" cellpadding="0" border="0" style="MARGIN: auto auto auto 9pt; 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="204" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 153pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">Materials</td><td valign="top" width="108" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 81pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">Quantity</td><td valign="top" width="360" rowspan="4" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 270pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;"> Note: The kit contains sufficient reagents for 250 assays performed in 96-well tissue culture plate.* b-Mercaptoethanol was not provided in the kit</td></tr><tr style="mso-yfti-irow: 1;"><td valign="top" width="204" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 153pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">Assay Buffer</td><td valign="top" width="108" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 81pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">20 mL</td></tr><tr style="mso-yfti-irow: 2;"><td valign="top" width="204" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 153pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">ONPG</td><td valign="top" width="108" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 81pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">4 mL</td></tr><tr style="mso-yfti-irow: 3; mso-yfti-lastrow: yes;"><td valign="top" width="204" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 153pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;"><a name="OLE_LINK2"></a><a name="OLE_LINK1">b</a>-Mercaptoethanol*</td><td valign="top" width="108" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 5.4pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 5.4pt; PADDING-BOTTOM: 0cm; BORDER-LEFT: #ece9d8; WIDTH: 81pt; PADDING-TOP: 0cm; BORDER-BOTTOM: #ece9d8; BACKGROUND-COLOR: transparent;">100 mL</td></tr></tbody></table>

macgene 发表于 2009-1-5 09:45:00

<p class="MsoNormal" align="left" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; LINE-HEIGHT: 125%; TEXT-ALIGN: left; mso-pagination: widow-orphan; mso-list: l0 level1 lfo1; tab-stops: list 18.0pt;">Ø       For each well, add <chmetcnv wst="on" unitname="l" sourcevalue="200" hasspace="False" negative="False" numbertype="1" tcsc="0">200 </chmetcnv>ml assay buffer and 5-10 ml of cell lysates.<p class="MsoNormal" align="left" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; LINE-HEIGHT: 125%; TEXT-ALIGN: left; mso-pagination: widow-orphan; mso-list: l0 level1 lfo1; tab-stops: list 18.0pt;">Ø       Incubation at 37 ºC for 20-60 minutes till yellow shade is observed.<p class="MsoNormal" align="left" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; LINE-HEIGHT: 125%; TEXT-ALIGN: left; mso-pagination: widow-orphan; mso-list: l0 level1 lfo1; tab-stops: list 18.0pt;">Ø       Add 100 ml stop solution (<chmetcnv wst="on" unitname="m" sourcevalue="1" hasspace="True" negative="False" numbertype="1" tcsc="0">1 M</chmetcnv> Na2CO3).<p class="MsoNormal" align="left" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; LINE-HEIGHT: 125%; TEXT-ALIGN: left; mso-pagination: widow-orphan; mso-list: l0 level1 lfo1; tab-stops: list 18.0pt;">Ø       Measure O.D. at 420 nm.<p class="MsoNormal" align="left" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; LINE-HEIGHT: 125%; TEXT-ALIGN: left; mso-pagination: widow-orphan; mso-list: l0 level1 lfo1; tab-stops: list 18.0pt;">Ø       Calculate the units of b-galactosidase activity using the formula: b-gal (unit) = (O.D.420 nm X 380 X 10) / (minutes of incubation X volume (ml) of cell extract)  Reference: Herbomel et al. Cell 39:653, 1984

macgene 发表于 2009-3-30 14:33:30

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[原创]β－半乳糖苷酶检测试剂盒