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Questions related to Blotting Techniques
We want to remove the high abundance proteins in plasma by using 'Pierce Top 12 Abundant Protein Depletion Spin Columns' so that we can use this plasma for 2D-DIGE. Thermo Fisher advises max 10 µl...
02 February 2016 1,315 2 View
Hi, I ran two gels yesterday (Beta Actin) and did not get good bands. I am very sure that it has nothing to do with my pipetting (I have ran the same samples before and got good bands), therefore...
02 February 2016 4,934 6 View
In my lab we have the Trans-blot Turbo by BioRad, and I wonder Pros and Cons of this technique against the conventional transfer. I guess the main pro is a quick transfer (~30') but i wanna know...
18 January 2016 7,500 1 View
01 January 2016 1,029 1 View
I need to transfer a membrane protein (size 82.9kDa) onto a PVDF membrane. Can anyone help me with the transfer conditions I need to use for this? Is 70V for 4 hrs enough or should I transfer even...
30 December 2015 2,026 2 View
12 December 2015 2,894 3 View
I keep trying Western Blotting using lysozyme antibody from SC and abcam, however I couldn't detect the band. Is there any suggestion that I can get through this? I used donkey anti-goat for a...
10 October 2015 2,818 3 View
After I obtained an SDS-PAGE of my purified protein with single band having molecular mass around 66 kDa, I decide to run my sample on a gel filtration column and calculated thatba 66 kDa Protein...
08 June 2015 8,882 7 View
06 June 2015 1,903 7 View
Since ELISA kits are very expensive, was wondering if I could measure that with western blot technique? and If yes, how to measure the protein concentration? For the cell lysate I use protease...
07 April 2015 2,586 7 View
04 April 2015 8,008 8 View
Blotting the same membrane for two different protein saves the time and cost. There are many ways of doing this either with stripping or without stripping. Which is the better approach and can we...
13 January 2015 430 2 View
01 January 2015 9,565 3 View
Hi all, I'm confused about some literature on the dot blot and Far-WB, since it is claimed that Far-WB can be used to detect interaction between prey and bait proteins but there is no such claim...
12 December 2014 9,060 4 View
Blotting Techniques
12 December 2014 2,793 1 View
I would like to store my blot after transferring, i'm just not sure how to do that. I have once tried to leave my blot in PBST over few days and I found that the signal I got after probing wasn't...
11 November 2014 5,100 2 View
I am working with phosphorylated proteins, using the western blotting technique. I have seen in the literature both phosphorylated proteins normalized with structural proteins, such as actin, or...
10 October 2014 4,110 10 View
I have been trying to use the reverse line blotting to detect several tick borne pathogens. However, I get very large signals. I have decreased the concentration of both PCR products and the...
10 October 2014 8,345 8 View
I am looking for a protocol to quantify proteins in the mice brain by western blot technique. I want to see the effect of treatment on AD pathology and neuroinflammation markers. I would like to...
09 September 2014 8,077 8 View
I am experiencing problems with one-dimensional Blue Native PAGE: It seems that proteins in the rage of 300 - 600 kDa are not properly transferred onto the PVDF membrane during Western Blotting of...
07 July 2014 8,570 6 View
05 July 2014 9,504 5 View
Western blot.
06 June 2014 2,866 1 View
I am trying to endogenous express MDR1 protein in a mammalian cell and I would like to confirm the presence of the expressed protein in the plasma membrane of different patches of cell populations...
01 January 2014 7,561 1 View
Did anybody ever test this?
01 January 2014 2,009 3 View