Arrhenius Lin

9 Questions 33 Answers 0 Followers

Questions related from Arrhenius Lin

How long can PhosSTOP protect phosphotyrosine from PTP dephosphorylation for?

IP is usually used to confirm protein phosphorylation, but if the antibody used has a weak affinity to the target protein, can we extend the incubation time to overnight if the cell lysate is...

01 January 2014 6,583 3 View

Why can't I induce autophagy in HeLa cell?

I have trouble inducing autophagy in HeLa Cell. HeLa cell was transfected with pmCherry-LC3B for 24 hours, and EBSS（with 0.1% w/v glucose) was used to induce autophagy for 2 hours.The IF image is...

10 October 2013 6,227 2 View

Are there any forms of cancer reported in invertebrates?

A lot of cancer researches have been focused on mammals, I am just curious whether this exists in invertebrates such as C.elegans, D. melanogaster etc

08 August 2013 5,467 9 View

Why can't I reproduce the result from my first in vitro phosphorylation assay?

I have tried to perform an in vitro phosphorylation assay by using a purified tyrosine kinase and its potential substrate.In the first try,I got a perfect result, but I can't replicate such...

08 August 2013 7,649 16 View

Can we deduce a 3D structure of a protein from its fragment whose 3D structure is known?

There are tens of thousands of 3D structure in PDB,and a portion of them are fragments but not full-length due to some flexible motif will adopt disordered conformation.The question is that, if we...

08 August 2013 2,137 5 View

Is there some transfection method that delivers plasmid in a endosomal-lysosomal independent pathway?

I have been trying to knockdown a gene of lysosome recently. When cells are transfected with pSuper-GFP-neo-target shRNA, the resulting transfectants seems hard to receive other plasmids for a...

05 May 2013 9,061 3 View

How to express large recombinant proteins with molecular weight over 100kDa in E.coli?

I have been trying to express some large eukaryotic proteins,over 100kDa in sizes, in BL21(DE3) and Rosetta(DE3), the outcome is frustrating.Does anybody have any ideas about how to achieve this goal?

03 March 2013 6,078 12 View

Can anyone suggest ways to optimize my co-transfection?

I have been working on transfecting HeLa cells with two expression vectors using Turbofect. One is pCI-Neo and the other is pEGFP. The transfection rate of pCI-Neo is quite low, however, in...

12 December 2012 5,052 21 View

How can I interpret this image?

I am a rookie in cell biology and I am performing an overexpression of my target protein in the HeLa cell. The expression vector I am using is pCI-neo. I have observed some puncta subcellular...

12 December 2012 8,320 24 View