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

Could someone suggest some methods for the purification of proteins from inclusion bodies?

As most of the proteins are going into the insoluble fraction as an aggregate and a very low amount in soluble fraction...

08 September 2013 579 5 View

12% ledeburitic cold work tool steel, etched for revealing retained austenite, any help for clarifying?

Hello everyone. Given microstructure image belongs to a 12% ledeburitic cold work tool steel (annealed and tempered). The sample was etched with picral (100 mL ethanol and 4 g picric acid). I...

02 March 2021 4,119 3 View

RIA. How to measure?

When you use RIA, with a control the unknown sample with antibodies, you bare in mind the binding sites of the antibodies. However you still need to measure labelled antigen (radioactive) and not...

28 February 2021 2,133 3 View

What is the most suitable resuspension buffer for TCA/acetone precipitated protein?

I need to extract protein from fermented carbon sources for Bradford assay. Most researchers experiencing insolubility of pellet in resuspension buffer. Please assist me to select most suitable...

26 February 2021 7,129 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

Standard curve shifting over time. How do I stabilise the ELISA?

24 February 2021 1,365 3 View

Steps to calculate activation energy by coats & redfern method?

I'm a chemistry student and I'm researching the kinetic degradation of mixed plastic waste. I'm having trouble calculating the activation energy by the coats & Redfern method. The data that...

24 February 2021 974 1 View

Is there an ideal incubation time of RGO-modified SPCE before subsequent use, i.e., electrochemical characterization or further modification?

I notice that upon washing and air-drying of my RGO/SPCE surface and characterizing again via DPV, the current response towards ferricyanide probe significantly decreases. My worry is that the...

22 February 2021 5,324 3 View

How to calculate the fraction metabolized of a drug by an enzyme using clearance(ml/min) values of the metabolite formed?

I am trying to find the fraction metabolized of a drug by CYP and UGT enzymes. I am unable to get the values of fraction metabolized (Fm) directly from the literature. Can we calculate the Fm of a...

21 February 2021 5,379 2 View

How can I measure the surface interaction or surface interfacial chemistry between polishing abrasives and upper surface of substrate ?

I am working on polishing abrasives. I need to do few characterizations ? which can provide the on time surface chemistry of substrate with polishing abrasive during optical polishing. I would...

18 February 2021 5,319 3 View

What is the proper in vitro culturing method to get information about the activation of innate immune cells by VLPs?

Dear All, I am interested in studying the response of immune cells against virus-like particles (VLPs). I used the PBMC in vitro assay and treat it with the VLP using PHA as a positive ctrl while...

16 February 2021 1,337 2 View