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

Does the pressure(-600 mmHg) affects pH?

When I conducted the degassing experiment using decompression under anaerobic condition(and I used ferrous based chelating agent), the oxidized ferric were observerd. So, could you tell me the...

03 March 2021 4,892 5 View

How to increase a model accuracy ?

dear community, my model is based feature extraction from non stationary signals using discrete Wavelet Transform and then using statistical features then machine learning classifiers in order to...

03 March 2021 6,994 5 View

No title

02 March 2021 3,060 3 View

New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before?

Question to you and THEM, the New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before? There appears to be a core problem for...

02 March 2021 3,024 2 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 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

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 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

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View