Recipe for protein extraction from coral tissue?

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Can we do different species of animal cells in 2 dimensional gel electrophoresis (2DE) protein separation?

For example I am going to extract protein of parasite inside the muscle of the fish (I cannot totally/fully isolate the parasite due to its small size from the muscle of the fish but I get the...

31 December 2018 6,857 2 View

How can I represent SDS-PAGE results for machine learning analysis?

I am trying to classify and analyze the results of an SDS-PAGE based array for bacterial detection using machine learning, but I have trouble finding the best way to represent the results with...

27 February 2021 9,176 3 View

Why my agarose gel staining sometimes looks not homogenic but like gradient (see the figure)?

I am worried about this overexposure of the upper part of the gel in the picture. Is it possible that this is the result of too much concentration of ethidium bromide? Why is there such a big...

25 February 2021 8,140 3 View

EMSA troubleshooting. Protein-DNA complex not migrating, cold probe lane with multiple bands. Can anyone help?

I'm running an EMSA to show the DNA binding activity of recombinant proteins versus the Wild type. Previous assays run by my research group using a different test system show the protein-DNA...

24 February 2021 8,325 1 View

Problem with ligating 2.8 kb insert into 8.5 kb plasmid?

I am trying to clone 2 copies of the same mammalian gene into the pSF-CMV-Ub-Puro Ascl (contains HindIII, KpnI and NheI cut sites) plasmid. This is what the final product is supposed to look like...

24 February 2021 6,310 3 View

How to manage western blot for proteomics data validation?

Hi Everyone, I am a bigginer in western blotting and I am trying to use it for validation of my proteomics data. I have: two regulated proteins with molecular mass of 40 and 73 kDa one control...

23 February 2021 6,671 4 View

Alginate and silk fibroin hydrogels toxic to cells?

Hello, I asm currently working with alginate and silk fibroin hydrogels. I prepared a blend of alginate and aqueous silk fibroin (straight from dialysis) and resorted to ionic crosslinking to...

23 February 2021 8,249 2 View

Diltuions for a Western blot?

I need help using the equation C1V1=C2V2 figuring out how much antibody to add to use for a western blot. The stock was origianlly 3.45 ng/ul but diluted 1:10 so now its 0.345 ng/ul. I need to run...

22 February 2021 9,901 3 View

Why the nucleic acid gel melted after running for 30min?

Hi all, I proceed with nucleic acid gel electrophoresis recently several times, but a wried phenomenon is they always melted after running over 30min. My gel concentration is 1% percentage...

22 February 2021 9,896 3 View

Sanger Sequencing results not matching experimental results of Restriction Digestion, colony PCR and protein expression?

I have cloned two domains of a gene into two bacterial vectors using cut and paste cloning with different RE sites. I checked the putative clones by colony PCR and Restriction Digestion (fallouts...

18 February 2021 6,736 4 View

What is the cause of the banding obtained after purifying the PCR product with ExoSAPIT?

Hello We are working on a PCR product with an approximate size between 5000 and 7000 bp, we want to use it as a positive internal control, since we have sequenced it and it corresponds to the...

17 February 2021 2,896 3 View

Abhijit Mitra

Downs, C.A. (2005) Cellular diagnostics and its application to aquatic and marine toxicology. In: G. Ostrander (ed) Techniques in aquatic toxicology. 2. CRC Press, Inc., Boca Raton, pp 301‐313 Ghosh, S., Gepstein, S., Heikkila, J.J., et al. (1988) Use of a scanning densitometer or an ELISA plate reader for measurement of nanogram amounts of protein in crude extracts from biological tissues. Analytical Biochemistry, 169, 227‐233. Rasband, W.S. (2011) ImageJ, U. S. National Institutes of Health, Bethesda, Maryland, USA, http://imagej.nih.gov/ij/, 1997‐2011. Sokal, R.R. and Rohlf, F.J. (1995) Linear Regression. In: Biometry the principles and practice of statistics in biological research, 3rd edn. pp. 452‐455. W.H. Freeman & Co., New York. -- can you consult these please?