Which detergent interferes with enzymatic activity the least?

I am doing an assay for interaction with a substrate and I need a detergent that will interfere with the enzymatic activity as little as possible. What is the general recommendation in this case....

10 November 2019 9,907 6 View

Is there a his-tagged protein standard for IMAC HPLC?

I am doing some his-tagged protein purification and I would like to see if the chromatography works by running a standard. Is there a cheap histidine tagged protein people tend to use? Also the...

03 April 2019 2,537 5 View

Is there an algorithm prediction for peptide absorbance at 214nm?

I have a peptide I would like to quantify with 214 nm abrobance. Is there an algorithm (or a rule of thumb,) to predict what the absorbance should be like there for 280nm?

03 April 2019 7,211 1 View

How to make filter tips from normal tips?

I need to get a filter of good porosity (flows well) at the bottom of a 200ul tip. I need to filter small volumes of solutions in 96 format. Any ideas? The filter needs to be inert (so C18 or C8...

31 December 2018 6,909 4 View

Please recommend a dye that can be quantified by absorption to inverstigate gel filtration desalting efficiency?

I would like to test the efficiency of desalting by a gel filtration column and I decided to check the desalting efficiency by monitoring removal of a dye through absorbance. Can anyone recommend...

01 February 2018 2,886 2 View

Can I treat protein sample as part of the water phase with chloroform-methanol protein precipitation?

03 April 2017 8,178 1 View

Offline fraction of peptides by HILIC using disposable columns?

I need to perform offline fractionation by HILIC. I have done offline fractionation using disposable RP columns (I have used home made columns with oligo R3 material, but Thermo sells disposable...

10 November 2016 4,175 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

Does the pressure(-600 mmHg) affects pH?

When I conducted the degassing experiment using decompression under anaerobic condition(and I used ferrous based chelating agent), the oxidized ferric were observerd. So, could you tell me the...

03 March 2021 4,892 5 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

Does any one can help me choose a reasonable basis set for intermediate metal ions interactions with guanine (i.e Fe2+, Mn2+, Cu2+ and exc.)?

Hello every body Does any one has any idea to help me choose a reasonable basis set for intermediate metal ions (i.e Fe2+, Mn2+, Cu2+ and exc.) interactions with guanine or other DNA organic...

01 March 2021 6,187 2 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

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

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

Cloudy/high background western blot - can anyone help me?

Hi everyone! I try to detect VDR protein (~48 kDa) using Western Blot. After SDS-PAGE using 10% MP TGX Stain-Free Gel from BioRad I transferred the protein onto an metOH activated PVDF membrane...

23 February 2021 6,266 3 View