How do I troubleshoot this solubility problem of PLGA nanoparticle loaded with protein and prepared by Double emulsion method?

05 June 2015 9,706 5 View

How can one use DIPEA for deprotonation of primary amine groups in polyethylenimine for protein conjugation?

31 December 2014 415 2 View

How do you determine the number of carboxyl groups in a Graphene oxide?

I wish to know the amount of carboxyl groups in graphene oxide for a conjugation experiment. Is there any technique like acid base titration? If so can anyone provide a protocol for that?

09 October 2014 698 3 View

How can I calculate the degree of thiolation of a material containing free thiol group using Ellman's test?

I have prepare thiolated PEI and I want to know the degree of thiolation. So I did the ellman's test with cysteine as the standard. Now how to calculate the degree of thiolation from this...

05 June 2014 1,423 1 View

Is a copper II acetate test meant for checking the primary and secondary amine groups in a polymer?

Ninhydrin is used for testing the primary, secondary, and tertiary amines of synthetic cationic polymer or protein. Is the copper II acetate test only meant for primary and secondary amines in the...

04 May 2014 3,452 1 View

How can I calculate the DNA bp: dye ratio?

I have YOYO-1 dye and I want to add dye at 5:1 ratio. And I am using a plasmid of length 5411 bp. Please tell me how to calculate this ratio to get a volume of dye to be added to DNA of 1 microgram.

01 February 2014 1,444 2 View

Can 2% SDS be used as a reaction buffer for Ellman's test?

For a Free Thiol assay, I wanted to used DTNB and a reaction buffer. But since some materials for making reaction buffer is not available. I wanted to use SDS as reaction buffer.

31 December 2013 3,900 1 View

RNA gel electrophoresis - denaturing or non-denaturing??

If I want to just look at the RNA integrity (of RNA stored in -80 for more than a year), should I go for denaturing or non-denaturing electrophoresis? I've seen most people use formaldehyde gels...

02 March 2013 6,224 16 View

Standard curve for relative real time PCR

I have a control cell sample and drug treated cell sample in my experiment. I want to know which sample to be taken for creating a standard curve in relative real time PCR? Should I use the...

02 March 2013 3,916 0 View

How to analyse agilent microarray data using MATLAB?

Using the 2012b version of MATLAB, I wish to know how to import the data and analyse the data for normalization, fold change analyses and background correction.

11 December 2012 9,519 1 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 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

Which fluorescent dye to use for measuring total ROS production?

Is DCFDA appropriate for measuring total ROS production, or can it only detect certain types of ROS? If I want to measure total ROS production, which dye should I use?

01 March 2021 9,447 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

Problem with ligating 2.8 kb insert into 8.5 kb plasmid?

I am trying to clone 2 copies of the same mammalian gene into the pSF-CMV-Ub-Puro Ascl (contains HindIII, KpnI and NheI cut sites) plasmid. This is what the final product is supposed to look like...

24 February 2021 6,310 3 View

Has anyone tried to scan fluorescence IHC slides with microarray scanner?

I have an old slide microarray scanner (VIDAR Revolution 4200) and would like to give it a "second life", as IHC scanner array (fluorescence dyes), FISH over tissue or similar. Has anyone tried...

24 February 2021 9,210 1 View

What is the proper range of FRET efficacy?

I would like to use FRET effect to probe the distance between two functional moieties (I am sure the distance is smaller than 10 nm). I have tried several times, but I found out that the FRET...

21 February 2021 3,562 1 View

Is nanotechnology part of materials science? Is it possible to obtain Pico technology and move from hypothesis to application?

Pico-scale This is three orders of magnitude smaller than a nanometer (and thus most nanotechnology) and two orders of magnitude smaller than most chemistry transformations and measurements. Pico...

18 February 2021 4,540 5 View

Does the property of GFP affect a fusion protein's localization?

Hello, I am planning to insert GOI between CaMV 35 promoter and mGFP in pCambia1302 to produce a fusion protein and see subcellular localization of the protein. But the vector says it's membrane...

15 February 2021 4,353 3 View

QPCR troubleshooting - what could cause an increasing ROX signal?

Hi everyone, I'm looking for some help troubleshooting a recent qPCR issue I've encountered. I am using fluorescent Taqman probes and a qPCR master mix that includes ROX as a passive reference....

08 February 2021 419 3 View