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Questions related to Blotting Techniques
Dear all, I have an issue figuring out good plunging conditions for large lipid structures: I have carbon coated C-Flat 2/2 400mesh copper grids with large (elongated) lipid structures on it....
14 December 2020 6,823 2 View
I used TEM equipped with tungsten gun to obtain attached image (several other too, like shown). Side blotting technique followed by negative staining (1% PTA for 30 sec) was used. I cannot get the...
01 November 2020 2,067 5 View
I would like to use my western blot membrane after re-probing (mild stripping) but I don’t know if will be able to use it often. Is there a way to store nitrocellulose membrane over the months...
02 February 2020 9,648 3 View
I have to detect dsRNA in bee larvae and pupae. I do not need to quantify. I would extract RNA, synthesize cDNA and try to amplify the target fragment by PCR. But in literature I have seen that...
01 January 2020 6,438 1 View
Hi, I want to carry out western blotting for a 233 kDa protein. I was told to use a tris-acetate gel due to the size of the protein. However, I was wandering would I be able to detect the...
26 March 2019 6,427 4 View
03 March 2019 4,572 5 View
I'm looking for a decent method to ship multiple strains of yeast on filter paper (around 85 mm in size) for shipping using liquid culture. I'm having little luck reviving yeast strains blotted...
10 October 2018 5,810 2 View
Well, I am really confused with this probe thing. For example, if I am looking for an unknown gene in a long stretch of DNA using Southern blot technique,how would I design a probe without knowing...
08 August 2018 9,037 0 View
the same procedure used for the DNA elution and purification. i need to know whether the nitrocellulose membrane carries any affinity towards DNA during the blotting like the silica matrix does...
08 August 2018 957 4 View
Dear all, I have been having problems with various antibodies in my western blots. For some reason they remain on the blot even after they were washed off (0.5% TBST 3x 5 minutes). I have tried...
03 March 2018 1,103 4 View
I'm looking to update our LAS4000 gel and western blot image system. We have a Licor but would like to keep chemiluminescence capabilities. Does any one have recommendations or suggestions for...
24 July 2017 9,745 11 View
Does anyone know of some reference protein antibodies that can be used for western blot to distinguish the amount of grey and white matter in your sample? Or do you have some other suggestion on...
14 July 2017 7,047 2 View
I did a western blot with a PVDF membrane. Because I only have 5 samples, I have thought of using 5 ml (1:1000 antibody dilution) of antibodies (Ni-NTA AP Conjugate). Unfortunately, I have...
12 July 2017 1,456 2 View
07 July 2017 6,753 3 View
07 July 2017 6,156 10 View
07 July 2017 625 2 View
We make our own gels for the western blots, both 7.5% and 10%. The top half of the gels appear to look fine when we are imaging them, but the bands on the bottom half are extremely fuzzy, light...
07 July 2017 1,476 2 View
I am very new to western blotting and I am not really sure about how to set up my western blot. I have a treatment and a control group, each with around 7 samples from the eyes of chicks. I want...
07 July 2017 938 4 View
I've tried lowering DNA quantity, TAq quantity, and primer concentration. I also tried adding Qiagen's ''Qsolution'' with absolutly no difference to my results. In comparison, my fungal ITS PCR...
07 July 2017 9,914 4 View
I am wondering If anyone have used dopamine receptor 1 (DrD1) and got clear bands after running in gel. I have used one which sometimes gives one band, sometimes give additional band near the...
07 July 2017 762 4 View
I have completed the imaging process using chemiluminescence method and would want to preserve the blot for future reference. Can I just wrap and seal the blot and save in -20 degrees? Any inputs...
07 July 2017 9,347 3 View
I am struggling to get my LC3-I and LC3-II bands to separate on my western blot. I can see there is a difference in the protein levels but I'm not getting any distinction between the two. I am...
28 June 2017 2,420 0 View
06 June 2017 6,660 1 View
Hi, I have trouble detecting a protein in a western blot. In my current blots I can’t see any bands at all. In my experiments I am looking at different proteins in unphosphorylated and...
23 May 2017 8,820 10 View