Can anyone report on the molecular weight of Q5 Polymerase?

We are repeatedly troubled with strange Protein bands in PAGE after PCR using Q5 polymerase from NEB. Who has measured the molecular weight of Q5 already and could help us out? Thank you all in...

31 December 2019 5,875 3 View

Who can help in collecting export files of fluorescence spectrometers?

I am writing a script for Processing fluorescence spectra. In Order to make it useful for a large audience, I am collecting Sample Data of Export Files from fluorescence spectrometers of different...

03 April 2018 7,945 11 View

New inducer molecule - which journal to publish?

Dear all,I was able to identify a new, synthetic molecule which is able to induce expression in bacterial gene regulation (only a particular operon is effected) - The practical use of this...

11 December 2016 7,073 0 View

Ion exchange chromatography strategy?

Dear all, after IMAC elution, my E. coli expressed protein (pI=6,32, MW=13 kDa) is in 400 mM Imidazol and 0,3 M KCl. TEV-Protease and cleaved off His-Tag are also swimming in this solution. I want...

05 June 2016 3,754 12 View

Strange Gel-Permeation Chromatogram?

Dear all, my latest gel permeation purification produced some strange results (see chromatogram attached).I am purifying a small 14 kDa protein after Ni-NTA-affinity chromatography and subsequent...

04 May 2016 1,063 7 View

Lysate way too viscous, impossible to apply to affinity-column?

Dear all, my E. coli lysate is extremely viscous, to the point that it is impossible to apply it to the column for purification. What can I do?I added 500 units of DNAse to 50 ml, but the...

04 May 2016 9,555 8 View

Problem with protein dimerization?

Dear all,I am trying to purify recombinant prion protein (PrP).The chromatogram during gel permeation chromatography shows that nearly half of my protein exists in the fom of dimers. Running...

04 May 2016 9,918 4 View

Fungal contamination of E. coli culture?

Dear all, lately I get some serious contamination in my E. coli cultures. I suppose it is a fungus of some kind. I am using minimal medum with no trace elements and no vitamins, the only organic...

04 May 2016 8,895 46 View

Ammonium sulfate precipitate does not sediment, but floats?

Dear all, I am using ammonium sulfate precipitation of protein as a first purification step from my crude bacterial lysate.The method works well, the proteins precipitate in sequential order as...

04 May 2016 3,536 9 View

Which amino-derivatized sugars are metabolized by E. coli?

Dear all, I wondered which amino-sugars can be digested by E. coli.Sure, Glucosamine can be metabolized by these bugs. But which other amino-derivatized carbohydrates can they devour?What about...

03 April 2016 4,326 4 View

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02 March 2021 3,060 3 View

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

What could be some reasons why unstimulated monocytes are getting activated?

Im doing PBMC isolation -> CD14+ enrichment using magnetic beads -> stimulation setup. My negative control is just cells in cRPMI but they seem to get activated over and over again.

28 February 2021 7,883 3 View

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28 February 2021 9,936 2 View

When is it unnecessary to change organisational culture?

As a former management practitioner, I recall a generally accepted view that changing organisational culture is difficult, so you should only attempt it if the organisational change that you are...

27 February 2021 2,595 8 View

Does RNAiMAX transfection reagent work in serum free medium?

I diluted siRNA and RNAiMAX in opti-MEM and added to the cells which they were in the growth medium. Is it a right way? or should I culture cells in the opti-MEM medium for a while and not in...

26 February 2021 10,041 3 View