Acc to Lowry's protocol we need to grind the sample in buffer. Can you please suggest me which buffer to select for protein estimation?

Vegonia Marboh @Vegonia_Marboh

09 September 2018 1 9K Report

Lowry's method

Devtulya Chander

i am using phosphate buffer for protease assay which involves folin reagent

https://www.sigmaaldrich.com/life-science/learning-center/life-science-video/universal-protease.html

check at what pH particular protein/s is/are dissolvable

and prepare buffer of that pH. The link below could come handy while making buffers

https://www.sigmaaldrich.com/life-science/core-bioreagents/biological-buffers/learning-center/buffer-reference-center.html

For compatibility of different buffers the below PDF may be a useful one

https://assets.thermofisher.com/TFS-Assets/LSG/Application-Notes/TR0068-Protein-assay-compatibility.pdf

Badges
Science method

More Vegonia Marboh's questions See All

What does it mean by free glucose in the digestibility assay of starch?

I want to analyse the in vitro digestibility assay of starch. I have downloaded many papers explaining the procedures of in vitro digestibility assay but the protocols were different from paper to...

06 July 2018 7,462 0 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

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

Measuring Total immunoglobulin whole-body of fish larvae?

Hi Dear All, I prepared the extract of fish larvae whole body with PBS. Then protein concentration of the whole-body extraction samples determined by the Folin-Ceiocalteu-Phenol method (Lowry...

17 February 2021 1,971 3 View

How to improve the buffer capacity of sea water?

Hello everyone I would like to improve the buffer capacity of seawater (pH ~8) in order to dissolve as much CO2 as possible without dropping the pH below 7.5 (a range between 7 - 7.5 is...

17 February 2021 2,797 2 View