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 JoVE Biology

Staining Proteins in Gels

1, 2

1Keck Graduate Institute of Applied Life Sciences, UVP, LLC, 2Proteomic Center, Keck Graduate Institute of Applied Life Sciences

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    Summary

    Following separation by electrophoretic methods, proteins in a gel can be detected by several staining methods. Staining of proteins with Coomassie Blue, Silver Staining, SYPRO Orange, SYPRO Ruby are demonstrated in this video.

    Date Published: 7/08/2008, Issue 17; doi: 10.3791/760

    Cite this Article

    Gallagher, S., Chakavarti, D. Staining Proteins in Gels. J. Vis. Exp. (17), e760, doi:10.3791/760 (2008).

    Abstract

    Following separation by electrophoretic methods, proteins in a gel can be detected by several staining methods. This unit describes protocols for detecting proteins by four popular methods. Coomassie blue staining is an easy and rapid method. Silver staining, while more time consuming, is considerably more sensitive and can thus be used to detect smaller amounts of protein. Fluorescent staining is a popular alternative to traditional staining procedures, mainly because it is more sensitive than Coomassie staining, and is often as sensitive as silver staining. Staining of proteins with SYPRO Orange and SYPRO Ruby are also demonstrated here.

    Protocol

    The complete text protocol for this experimental approach is available in Current Protocols in Molecular Biology.

    Disclosures

    The authors have nothing to disclose.

    Comments

    8 Comments

    WHICH REACTION EQUATION OF CONVERT Cu+² TO Cu+1 in the protein test
    Reply

    Posted by: AnonymousAugust 3, 2008, 7:45 AM

    thank you!very good!
    Reply

    Posted by: AnonymousSeptember 6, 2008, 8:51 PM

    It is very good, thanks
    Reply

    Posted by: AnonymousNovember 12, 2008, 6:43 AM

    A good and cheap alternative to SYPRO Rubi is RuBPS staining. Here are some protocols. More information can be found on www.ruthenium.ag.vu   RuBPS staining protocol I (quality) 1. Fix the gel in 30% EtOH, 10% acetic acid overnight ². Rinse the gel in ²0% EtOH for 30 min and repeat 3 times 3. Incubate the gel in 1 mM RuBPS solution for 6 h 4. Equilibrate the gel in water for 10 min and repeat once 5. Destain the gel with 40% EtOH/10% acetic acid for 15 h 6. Equilibrate the gel in water for 10 min repeat once and scan   all% are in V/V   Procedure as published in Proteomics ²004, 4, 599–608.     RuBPS staining protocol II (fast)   1. Incubate the gel in 50 ml of 40% Ethanol/10% acetic acid containing    1 mM RuBPS for 1 h. ². Destain the gel for ²0 min in 40% Ethanol/10% acetic acid 3. Wash the gel for 10 min in water and scan   all% are in V/V   Procedure as published in PLoSONE. ²007 Feb ²8;²(²)e²63.       RuBPS staining protocol III (co-electrophoretical)   1. Add 1 ml of ²0 mM stock solution to one pocket of your SDS gel. Run     the gel according to your standard procedure. ². Incubate the gel in 50 ml of 40% Ethanol/10% acetic acid for ²0 min 3. Incubate the gel in 50 ml water for 10 min and scan       RuBPS staining protocol IV (loading buffer)   1. Add 1 ml of ²0 mM stock solution to your loading buffer (omit all other     dyes like bromophenol blue). Run the gel according to your standard     procedure. ². Incubate the gel in 50 ml of 40% Ethanol/10% acetic acid for ²0 min 3. Incubate the gel in 50 ml water for 10 min and scan  
    Reply

    Posted by: AnonymousFebruary 16, 2009, 1:20 PM

    Itz realy ² gud!
    Reply

    Posted by: sathya r.May 18, 2009, 11:01 AM

    Thank yoy
    Reply

    Posted by: JASSIM A.May 20, 2009, 9:39 AM

    thank you
    Reply

    Posted by: ibrahim b.August 27, 2010, 12:31 AM

    thanks! This is very useful for my job!
    Reply

    Posted by: AnonymousAugust 31, 2010, 4:33 AM

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