Why is there cell debris are coming after 2nd or 3rd passage of the cells???

Our lab is facing a difficult problem for almost a year. After thawing the cells are okay, but after 2nd or 3rd passage there are debris present between the cells. We use mycoplasma removed cells...

17 January 2021 5,629 6 View

Can you help me for generating chicken swarm optimization matlab code?

Multi-objective optimization through chicken swarm optimization technique. Both maximization and minimization problem.

31 December 2020 2,299 4 View

Any good methods for double-stained markers quantifications from tissue sample using computer programs?

How can we best quantify double-stained markers when comparing diseased and control? For example, if we want to stain one marker of smooth muscle cells and compare the expression in diseased and...

21 December 2020 7,389 1 View

Membrane protein Expression Purification: Broad Asymmetric peak on size exclusion ?

Hello, I am expressing an integral membrane protein (MW 51kDa) in Hi5 insect cells. I initially purified the protein by Nickel-His, concentrated it and subjected it to Size Exclusion...

10 December 2020 2,781 5 View

How should I store the milk sample in RNAlater?

I want to extract RNA from human breast milk. How should I store the milk sample in RNAlater? And after RNA extraction how can I store the RNA?

01 December 2020 7,183 4 View

Can U suggest standard protocols of quantifying total phenolics & total flavonoids using gallic or tannic acid for the former and quercetin for later?

The problem is that everytime getting the flavonoids content much higher than phenolics which may be, but in rare occasion. In phenolics, I am adding100 ul sample with 900 ul dH2O with...

29 September 2020 4,993 6 View

What is the ideal Elution buffer composition for a His tag protein purification to prevent protein aggregation after freeze thaw ?

Hi All, Recently, I purified 8 proteins and stored at -20 with the elution buffer containing 20 mM Tris, 250 mM NaCl and 200 mM Imidazole. I autoclaved the buffer, so I do not know whether the...

25 September 2020 9,814 6 View

What is the use of having multi-wavelength absorption measurements in Aethalometer?

Aethalometers are used to estimate the light-absorbing aerosols or Black Carbon (BC) or Soot particles. There are widely used aethalometers which are measuring light absorption in multiple...

06 September 2020 1,050 3 View

Has anyone done magnetic target localization using Genetic Algorithm?

I am trying to model magnetic target localization in MATLAB using those algorithms. So essentially what I want to output is the position vector of the target (x, y, z) and the magnetic moment...

28 August 2020 9,811 2 View

Can an isolated protein domain give a single band in SDS-PAGE equivalent to its dimeric mol. wt. ?

The protein I'm working with forms a homodimer and is a 2-domain protein. I have cloned and purified one of its domains. However, when I run the domain on SDS-PAGE, I get a single band equivalent...

19 August 2020 7,893 14 View

How do I determine the degree of polymerization of PEG 400 based on NMR integration?

Hello, I am working on the photocatalytic degradation of PEG 400. I would like to know if it's possible to determine the degree on polymerization based on NMR integration, or by the MW of the...

02 March 2021 6,186 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

How to make fractions of chloroform extract of plant, using seperating funnel?

28 February 2021 8,466 2 View

How to include original NMR data in a manuscript?

hello. i am junior researcher. I received a comment from a reviewer that he could not find NMR data in my submitted paper. I had included the NMR graph with explanation and formula to calculate...

27 February 2021 734 4 View

How to manage western blot for proteomics data validation?

Hi Everyone, I am a bigginer in western blotting and I am trying to use it for validation of my proteomics data. I have: two regulated proteins with molecular mass of 40 and 73 kDa one control...

23 February 2021 6,671 4 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 to convert 2D stack images to 3D to obtain surface area of nucleus?

I have 15 slices of the nucleus at different z-heights from confocal imaging. I would like to use these slices to reconstruct the 3D nucleus and ultimately estimate the surface area of the...

20 February 2021 6,967 1 View

I can detect my protein but the encoding DNA sequence has stop codons within. Should I ignore the stop codons?

I constructed a synthetic DNA library (scFv, VH-VL orientation) with a 3' reporter and 8x histag. I cloned and expressed this gene following which I performed an ELISA. The ELISA results suggested...

19 February 2021 7,006 5 View

Is my GFAP staining in right position?

I was differentiate hESC to astrocyte. In the second week of differentiation, I stained my cell with s100beta and GFAP. I expected that the staining site of GFAP would come from the cytoplasm,...

15 February 2021 8,982 1 View

(western blot) can methanol damage my antibody signal?

I accidentally rinsed with methanol after adding my primary antibody to my membrane. I noticed after ~10 seconds and immediately replaced it with PBST (3x15 min washes). Will my antibody signal be...

12 February 2021 5,824 3 View