प्रोटीन के electrophoretic पृथक्करण


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इस वीडियो में, हम पाली acrylimide जेल वैद्युतकणसंचलन (पृष्ठ) का उपयोग प्रोटीन की electrophoretic जुदाई के लिए एक विधि का प्रदर्शन.

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Chakavarti, B., Chakavarti, D. Electrophoretic Separation of Proteins. J. Vis. Exp. (16), e758, doi:10.3791/758 (2008).

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वैद्युतकणसंचलन प्रोटीन का जटिल मिश्रण (जैसे, कोशिकाओं, subcellular भिन्न, स्तंभ भिन्न, या immunoprecipitates से) अलग, सबयूनिट रचनाओं की जांच, और प्रोटीन के नमूने की समरूपता को सत्यापित करने के लिए प्रयोग किया जाता है. यह भी आगे के अनुप्रयोगों में प्रयोग के लिए प्रोटीन शुद्ध सेवा कर सकते हैं. Polyacrylamide जेल वैद्युतकणसंचलन में, प्रोटीन एक polyacrylamide जेल मैट्रिक्स में एक pores के माध्यम से बिजली के क्षेत्र के जवाब में विस्थापित, रोमकूप आकार acrylamide एकाग्रता बढ़ाने के साथ घट जाती है. ध्यान में लीन होना आकार और प्रोटीन प्रभारी, आकार और आकार के संयोजन प्रोटीन के प्रवास की दर निर्धारित करता है. इस इकाई में, मानक Laemmli विधि असंतत जेल वैद्युतकणसंचलन के लिए denaturing शर्तों के तहत वर्णित है, सोडियम dodecyl सल्फेट (एसडीएस) की उपस्थिति में यानी,.


इस प्रयोगात्मक दृष्टिकोण के लिए पूर्ण पाठ प्रोटोकॉल में उपलब्ध है आण्विक जीवविज्ञान में वर्तमान प्रोटोकॉल .

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  1. Hi nice demo but I did not understand why the presenters kept on showing an agarose gel electrophoresed DNA gel in between as a representation instead of a protein gel.

    Posted by: Anonymous
    June 15, 2008 - 1:11 PM
  2. SYPROorange and SYPROruby staining - two fluorescent-based protein detection methods - can appear like ethidium bromide stained DNA gels.  So those gels you are speaking of may actually be protein gels. 

    Posted by: Anonymous
    June 16, 2008 - 9:34 AM
  3. Yeah possible,  but then why do we see only a single band.I guess SYPRO detection methods would detect the entire protein panel and not specific bands. unless otherwise they are detecting some specific enzyme activity using flourescent detection methods.

    Posted by: Anonymous
    June 16, 2008 - 9:52 AM
  4. We loaded BSA.


    Posted by: Anonymous
    June 17, 2008 - 2:12 AM
  5. Please not that we showed polyacrylamide gel electrophoresis and not agarose gel electrophoresis.


    Bulbul Chakravarti

    Posted by: Anonymous
    June 17, 2008 - 2:08 AM
  6. why cannot i see the video...? pls help me..

    Posted by: Anonymous
    October 10, 2008 - 3:35 AM
  7. i can see it now..thanks...

    Posted by: Anonymous
    October 10, 2008 - 4:50 AM
  8. my network is poor, I can not enjoy the video

    Posted by: Huang C.
    March 15, 2009 - 6:30 AM
  9. Hello, Please shoot me an email at nikitab@jove.com and we'll figure something out for you. Cheers, Nikita

    Posted by: Anonymous
    March 22, 2009 - 10:26 PM
  10. Hi, thanks what can I download your video ? please help me

    Posted by: violet s.
    January 14, 2012 - 10:35 AM

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