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Invitrogen NuPAGE Sample buffer


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#1 new2science

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Posted 03 May 2010 - 06:06 PM

Hi,

I am using a commerical gel electrophoresis kit from Invitrogen that contains pre-made gels composed of 4-12% Bis-Tris gel and along with the gel comes a kit to to prepare my samples before loading then into wells. To prepare the samples you add x amount of sample, then add the following to your samples: NuPage LDS sample buffer (4x), NuPage sample reducing agent (10x), and deionized water. I want to detect actin (because it is an ubquitous protein found in all cell types) from my cell lysates just to make sure my transfections worked properly. To lyse my cells I used M-PER and added protease inhibitor....so my question is I don't need to add any SDS loading buffer dye to my cell lysate beforer I prepare my samples using the sample preparation kit? I say this because the LDS sample buffer will do the trick to see the protein bands I'm interested in- is this correct????

Thanks

#2 new2science

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Posted 03 May 2010 - 07:11 PM

Hi,

I am using a commerical gel electrophoresis kit from Invitrogen that contains pre-made gels composed of 4-12% Bis-Tris gel and along with the gel comes a kit to to prepare my samples before loading then into wells. To prepare the samples you add x amount of sample, then add the following to your samples: NuPage LDS sample buffer (4x), NuPage sample reducing agent (10x), and deionized water. I want to detect actin (because it is an ubquitous protein found in all cell types) from my cell lysates just to make sure my transfections worked properly. To lyse my cells I used M-PER and added protease inhibitor....so my question is I don't need to add any SDS loading buffer dye to my cell lysate beforer I prepare my samples using the sample preparation kit? I say this because the LDS sample buffer will do the trick to see the protein bands I'm interested in- is this correct????

Thanks


Ah, nevermind...diregard this message. I figured it out I don't need to add SDS loading buffer to my cell lysate as the LDS contains a dye for visualization.




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