If their is no fragmentation of DNA in cancerous cell lines, does it proves their has been no apoptosis?

Where as in case of MTT assay, the results were positive, but in DNA fragmentation assay, their is no fragmentation of genomic DNA can be seen. So, my question is, is the DNA fragmentation solely...

07 August 2019 8,058 7 View

Which type of column should i use to check the purity of low molecular weight protein using FPLC reverse phase column chromatography?

03 April 2018 5,925 4 View

How to extract highly soluble low molecular wt. protein?

the supernant contains all the low molecular proteins dissolved, tried it with ammonium sulfate ppt, till it reached saturation point, even after that the protein can't be extracted. please suggest.

31 December 2017 2,345 0 View

In ammonium sulfate ppt, in order to check protein's activity, the precipitated pellet is not giving any zone of inhibition. why?

I have checked for activity with 30%, 60%, 80% precipitated pellet suspened in buffer, it didn't show any zone, whereas the treated supernatant such as 30%, 60%, 80% keep on showing zone against...

11 December 2017 9,265 0 View

What is the best fixative agent can be used for low molecular weight protein?

In SDS PAGE.,before staining procedure to be carried out, which one among the solution will be best for protein fixation? (low molecular weight 2kDa to 12kDa) 50% MeOH 50% H2O 25% Acetic acid...

09 October 2017 8,131 3 View

How to store gels for future reference?

After the gel has been properly destained, can it be stored for longer period? How? Kindly Tell me the storage buffer composition. Can we just pause the destaining process and, keep it in some...

09 October 2017 978 0 View

To plot a standard graph, what is the highest concentration of BSA that can be used?

As the unknown O.D i am getting is, 0.559, 1.324, 3.472 etc. what BSA concentration can be used to get a standard graph?

08 September 2017 2,797 1 View

In the first step of protein purificatio,does the protein degrades?the crude protein showed zone of inhibition but the 80%precipitated protein didn't?

why it is not showing zone? for how long the protein remain active after , precipitation, if kept in buffer?

01 January 1970 6,444 2 View

SDS Page - 10, 12, and 15%. Which will give better resolution for low molecular weight protein?

I have run 4-12% gel and i got this. I have tried with every gel percentage, still not getting anything,instead like this I got. Can somebody suggest why I am geting such thing...

01 January 1970 2,000 17 View

How can we choose buffer , for protein extraction/purification?

for low molecular, sensitive, generally shows activity below pH 5. for this kind of peptide(made up of few amino acids), which buffer will be the best? and how to choose? In case of ammonium...

01 January 1970 9,359 2 View

Protein-aptamer gel electrophoresis help, please??

Hi, I have problems with running gel electrophoresis. I have tried agarose gel electrophoresis and native PAGE. I have two proteins, which have molecular weights of ~30kDa and ~180kDa and two...

03 March 2021 4,275 4 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

What are some important techniques for protein induction and SDS-PAGE analysis?

Hello everyone. I am fairly new in protein chemistry field. I am trying to perform a protein induction hoping to purify my protein of interest using FPLC method. However, I am having trouble on...

03 March 2021 8,237 3 View

SDS page software for analysis?

I did sds page to determine the difference in protein expression between 3 types of vaccine , but i don't have scanner or densitometer available. . so i wonder if there is a software to analyze...

02 March 2021 6,270 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

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

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