Yu:Western: Difference between revisions
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(New page: =Western Blotting= {{Template:Yu}} __NOTOC__ ===Cell Lysis=== ===SDS PAGE=== ===Transfer=== ===Immunoblotting=== ===Developing===) |
Michaelcyu (talk | contribs) (sds-page) |
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=Western Blotting= | =Western Blotting= | ||
{{Template:Yu}} | {{Template:Yu}} | ||
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===SDS PAGE=== | ===SDS PAGE=== | ||
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|width="800px" valign="top"|1.[[Yu:SDS_PAGE|Cast SDS-PAGE gel]] and assemble the gel apparatus. | |||
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|2. Add 10μL of desired sample and 5μL 3x [[Yu:SDS PAGE Sample Buffer|Gel loading buffer]] to an eppendorf tube and boil for 5 minutes. Do this in separate tubes for each sample to be analyzed. After boiling, samples may be kept at room temperature until loaded | |||
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|3. Load 12μL to 15μL of sample into each lane. Load protein prestain marker, as well as BenchMark, if desired. Load any empty lanes with 1x [[Yu:SDS PAGE Sample Buffer|Gel loading buffer]] | |||
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|4. [[Yu:SDS PAGE#Running Gel|Run gel]] until the loading buffer is just running off the bottom of the gel | |||
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===Transfer=== | ===Transfer=== | ||
Revision as of 10:58, 14 June 2009
Western Blotting
SDS PAGE
| 1.Cast SDS-PAGE gel and assemble the gel apparatus. |
| 2. Add 10μL of desired sample and 5μL 3x Gel loading buffer to an eppendorf tube and boil for 5 minutes. Do this in separate tubes for each sample to be analyzed. After boiling, samples may be kept at room temperature until loaded |
| 3. Load 12μL to 15μL of sample into each lane. Load protein prestain marker, as well as BenchMark, if desired. Load any empty lanes with 1x Gel loading buffer |
| 4. Run gel until the loading buffer is just running off the bottom of the gel |
