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

Is there a way to separate free dye from fluorescently Labelled DNA?

I need to conduct a FRET analysis with labelled plasmid DNA and I am using YOYO-1 dye. I need to remove the free dye from the solution and get only the labelled DNA. Is there any technique with...

31 December 2013 8,505 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

Buffer to disrupt platelet alpha granules for PF4 quantification?

Hello! I want to quantify by ELISA the secreted (from platelets poor plasma) and the non-secreted (from platelet lysate) PF4 before and after TRAP stimulation. I will use the ELISA from R&D...

03 March 2021 1,499 2 View

Buffer Exchange using column packed with sephadex G-25 effect on protein concentration?

Hello, If i am doing a buffer exchange for an antibody of 1mg/mL, does the elution lose protein in the process of buffer exchange? For example, if i flow through 500 uL of 1mg/mL sample, and...

03 March 2021 6,299 3 View

How optimize the Size Exclusion Chromatography?

Hello! I'll do a size exclusion chromatography, but I only have an open column, and I'll perform the peptide extraction from yeast, using buffer lysis (sodium phosphate 50 mM/NaCl 30 mM/DNAse and...

01 March 2021 2,215 2 View

Why are my bands weird in this Agarose gel?

Can someone please give me some possible things that could go wrong? Here is my recipe: 0.5g Agarose 50 mL of TAE 1x 1 uL ethyl bromide. Gel was run at 100V for 1 hour. The buffer used is also TAE.

01 March 2021 9,952 3 View

What do I do with DLS peaks that are from the buffer?

I'm looking at the aggregation of my protein sample using DLS. Unfortunately, my buffer (20mM HEPES) also results in a set of peaks. These are at approximately 1 and 1000 d.nm. The lowest peak...

01 March 2021 9,015 2 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 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

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

What is a good reagent for lentiviral plasmid transfection in HeLa cells?

I have been using Lipofectamine 2000 but was hoping there may be a better protocol or one for something like Lipofectamine PLUS reagent.

24 February 2021 1,633 4 View

What urea buffer can i use for biotagged protein extraction from streptavidin beads?

Hi all, I have been doing research on biotagged LDB1 proteins in Mel cells. I purify the proteins using M280 streptavidin beads. The yield is very low so I've been trying to optimize it by...

24 February 2021 5,749 3 View