Which methodologies are the most appropriate for the determination of the activity of the enzymes related to the Nitrogen metabolism?

More Carlos E. Aucique-Pérez's questions See All

Anybody with an idea on how do design specific primers for detecting tomato resistance genes tm-1, TM-2 and Tm2^2?

I am trying to identify these 3 genes among some tomato cultivar collections and after aligning some sequences from NCBI, I couldn't find unique sequences to target for specific primers. There...

28 February 2021 606 3 View

Can I use acetic acid with an ELITE WAX Perkin Elmer column for Gas Chromatography?

Hi! I would want to know if I can use acetic acid with an ELITE WAX Perkin Elmer column for Gas Chromatography. This column has a polyethylene glycol phase, but I can't find the manual to check...

22 February 2021 730 2 View

Where can I find a reliable and updated methodology related with the design of spiral heat exchangers?

I need to design an spiral heat exchanger, buy I don't know what reference or bibliography (preferably a book) should I consult in order to get an updated and reliable methodology related with...

14 February 2021 7,910 3 View

Which are the factors that determine in a tourist the destination for their travel?

I'm working on a study of how selection criteria of Tourism have changed considering Pandemic + Vaccination expectation, so propose a new prediction behavioral dynamic model for Riviera Maya (Mexico).

09 February 2021 314 7 View

RWP1 growing in RPMI medium?

Hello everyone, Has anyone try to culture the normal prostate RWPE-1 cell line in RPMI medium supplemented with foetal bovine serum? There's some articles available but neither describe details...

08 February 2021 6,455 1 View

Can anyone recommend me a NMR Handbook?

Hi, I've been searching for a NMR Handbook with 29Si chemical shifts information but I can't find one. Also I've tried to find an online database with that kind of info but I have no lucky. Can...

02 February 2021 307 3 View

Equation to calculate Chlorophylls and carotenoids concentrations from a MeOH:HCl 1% (v/v) plant extract solution?

Hello everybody! I have performed an acidic MeOH:HCl 1% (v/v) plant extraction from Arabidopsis leaves to calculate Anthocyanins concentrations based on: Rabino, I., and Mancinelli, A. L. 1986....

25 January 2021 3,686 2 View

Cronbach's alpha or McDonald's omega? Which would be more correct and in what cases?

Cronbach's alpha or McDonald's omega? When it comes to reliability, we find different methods for its estimation. Immediately, the name of Cronbach's Alpha coefficient arises, as it is the most...

21 January 2021 330 4 View

Which aqueous solution should I test to make positive NPs stable to get mayor zeta potential +40 mV?

Hi, I need to find the correct media to make my positive charge NPs stable in an aqueous media. I have tried different solutions at different pH but the zeta potential does not increase more than...

21 January 2021 6,490 6 View

Mysterious line across SDS gel?

Does anyone have the similar issue? A line across the denature SDS gel shows up after staining with Coomassie blue and there are many lines in every lane if you take a closer look. This gel had 15...

07 January 2021 5,610 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

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

Can two EC50 values with different hill slopes be compared?

Hello, I fit a classical dose response curve Y=Bottom + (Top-Bottom)/(1+10^((LogEC-X)*HillSlope)) to my data (not related to inhibitors etc). The model I use is purely qualitative but it fits...

23 February 2021 8,155 5 View

Will SARS-CoV-2 Spike be broken into S1 and S2 by denaturing conditions?

Hello, I am performing a western blot of my SARS-CoV-2 Spike-pseudotyped lentiviral particles. I denature my virus by adding concentrated virus to 2X Laemmli buffer at 1:1, then boiling for...

17 February 2021 4,844 4 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

Why does flow deformation occur at the test line and control line on nitrocellulose membranes (LFIA)?

I use 1% BSA in 10 mM phosphate buffer (pH 7.4) solution when striping my nitrocellulose membranes with antigens and antibodies. However, I always observe deformed flow when my sample or running...

16 February 2021 4,224 3 View

What is the best temperature to coat an ELISA plate?

Hello, I want to control the temperature of my ELISA plate during incubations to get consistent results. In general, what is the best temperature to coat the plate with antibody? I have been...

16 February 2021 2,377 7 View

Can the viability assays (ex: MTT/CCK-8) measure cell proliferation? or are they solely viability assays?

I want to conduct a proliferation assay to test a chemotherapy combination on a cancer cell line. At first, I thought about the MTT assay. But now, all sources are saying that these do not measure...

14 February 2021 3,142 7 View

Ahmad Ali

GS can be assayed spectrophotometrically. Rest other enzymes can be measured by HPLC like techniques.

You can refer the following paper for protocols for nitrate assimilating enzymes

Ali, A., Jha, P., Singh, K. S., Raghuram, N. 2008. Spirulina nitrate-assimilating

enzymes (NR, NiR, GS) have higher specific activities and are more stable than those of rice. Physiol. Mol. Biol. Plants. 14(3): 179-182.

Carlos E. Aucique-Pérez

Dear Ahmad, thanks for your information.