How to determine the amount of acid required to stop the enzyme reaction?

I have to stop my enzyme reaction pH(8.5) by perchloric acid and further neutralized by KHCO3. How to determine the amount of acid that is required to stop the reaction and is there any...

05 June 2017 746 5 View

How to calculate the slope of a line that resembles the exponential decay?

Iam measuring the absorption of substrate in a enzymatic reaction to find out the kinetic parameters Km and Vmax. Plot of the curve (absorbance vs time) obtained in a exponential decay manner. To...

11 December 2016 8,770 3 View

How can I dissolve ZnCl2 in water?

04 May 2015 4,793 8 View

Is it possible to crystallize proteins which precipitate immediately after purification?

I have read in one of the article that they have crystallized protein by screening the protein solution which was partially aggregated before even setting and the drop and finally they got good...

03 April 2015 5,861 8 View

How might one overcome protein precipitation during elution ?

10 November 2014 8,925 13 View

Anyone familiar with the fate of expressed protein in E.coli?

If I have expressed a functional protein in e.coli,how the activity of the protein in bacterial cytoplasm will effect its survival? If it is a metabolic enzyme and by chance it is involved in...

04 May 2014 9,651 2 View

How can I eliminate the interference of sarcosine in the binding of protein to ni-nta?

I have solubilized my protein with 0.3% sarcosine and purified by using Ni-NTA,during purification most protein is going into flow through. I have diluted my sonicated sample to 0.1% sarcosine...

04 May 2014 8,160 5 View

What is the minimum percentage of sarkosyl that is allowed for a lysis buffer when extracting native proteins from inclusion bodies?

I have sonicated my bacterial cell pellet with 0.5% sarkosyl and have a sufficient amount in soluble fraction. In another condition, I have used 50mM each of L-Arginine and glycyl glycine in...

03 April 2014 2,923 3 View

I have purified my protein coupled to gst tag and i got several non specific bands during purification, how to reduce non-specificity ?

My protein has 59 kDa and I got gst along with it during purification. I am confused whether my protein is degrading or my induced colony has both native and/or vector with insert.

11 December 2013 8,488 1 View

Is it possible to use E.coli BL21(DE3) as a host for the expression of my target protein cloned in pQE 30 vector?

I have read some articles, in which they have used BL21 and in other some cases it is M15 cells. So, which is the better host strain for expression and why ?

08 September 2013 5,430 9 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

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

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

What is the most suitable server for predicting protein structures in bacteria?

I have used the i-Tasser several times, however it has been unavailable for several days. I tried the swiss-model, but the output was not very pleasant due to the model used. Is there any other...

28 February 2021 4,521 3 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

Are protein samples in SDS compatible with ELISA?

I have used an AllPrep DNA/RNA/Protein Mini Kit QIAGEN kit to extract RNA and protein from my samples. At the end of the protein purification, I resuspend my protein in 5% SDS. Will these samples...

28 February 2021 7,370 3 View

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View

Unknown debris sheets observed in cell culture flasks?

Hi, We culture neural progenitor cells in T175 culture flasks coated with PLO-Laminin. We have successfully grown these cells with our protocol for months. Recently, we observed large sheets...

25 February 2021 3,443 4 View

How to replicate the well conditions of a crystal hit with 20%(w/v) isopropanol?

I had a crystal hit in a well with 0.1M HEPEs pH 7.5, 10%(w/v) PEG 4000, and 20%(w/v) isopropanol. Why is isopropanol in w/v here if it's just solvent? How do I recreate this well condition?

25 February 2021 9,761 1 View