What transfer conditions should I use to blot a membrane protein of size 82kDa?

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I am doing alanine scanning mutagenesis using Pfu Ultra turbo DNA polymerase and XL1 blue supercompetent cells. My primers have the 3' overhangs. The annealing temperature I have used is 3C lower...

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What are the membrane transfer conditions I should use during western blot if i have two proteins of varying sizes?

If i need to do western blot of two proteins of varying sizes separated in the same gel (one 55kDa and one 13kDa), what should be the membrane transfer conditions that I need to use? Is 70V for 2...

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Does the use of Phos-Tag give a high background?

I will be incorporating Phos Tag in my protein gels to study histidine phosphorelay between two bacterial proteins. I wanted to know if there is a possibility of getting a high background upon...

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How can I solve my protein instability so it can be used in NMR and ITC studies?

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Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

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New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before?

Question to you and THEM, the New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before? There appears to be a core problem for...

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Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

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POlymer+DMSO solution and moisture??

What could be the reason of moisture at the sides of Vial when I am preparing my polmyer+DMSO solution for membrane fabrication? What etiquettes I am possibly missing here?

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Will a 10xHis-tag prevent anti-MBP from binding to MBP?

I have a protein with a 10xHis-MBP tag on the N-terminus. I'm fairly certain I am expressing my protein, but I'm not getting a signal on my western blot. Is there a possibility that the 10xHis tag...

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Unknown debris sheets observed in cell culture flasks?

Hi, We culture neural progenitor cells in T175 culture flasks coated with PLO-Laminin. We have successfully grown these cells with our protocol for months. Recently, we observed large sheets...

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What's the concentration recommend for recombinant protein use in western blotting?

I'm currently working having problem with the concentration for recombinant protein use in western blot, the one been used is Recombinant Human CD19 Fc Chimera Protein-rndsystem which does not...

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IF troubleshooting: looking at an intracellular protein that localizes to the cell's and how to reduce increase in signal due to cell ruffles?

I am currently trying to stain for PI3KBeta in MDA-MB-231 cells. My current issue is that 1) beta stains all over the cell and 2) there is an increase in signal at the edge of the cell and this...

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Can we recognize signs of the "volatility clustering" property of a time series in its time domain plot?

My purpose is to recognize the volatility clustering property without statistical tests such as The Lagrange Multiplier (LM) test.

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What is the best way to manage Western blots?

Increasingly, more articles are being retracted because of duplicate or otherwise dubious Western blots. Consequently, journals are asking for original Western blots and authentication of...

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