In vivo cross linking of mycobacterial membrane proteins?

I am trying to purify a membrane complex from mycobacteria. It seems that the interaction is very weak within the complex so I am not able to purify the whole complex. I am thinking to fix the...

08 September 2016 9,819 2 View

Why avidin is required for strep tag purification?

I am working with purification of protein by strep tag. In my protocol there is avidin in the binding buffer. Right now I do not have it. Can I proceed with the purification without adding avidin....

07 August 2016 1,053 3 View

How is GFP plus different compared to GFP?

06 July 2016 3,035 2 View

The concentration range of histidine for elution of proteins bound to Ni-NTA.

06 July 2016 9,566 6 View

How to isolate the quinol pool from the membranes?

I am interested in isolation of total quinols from the membranes. I want to use it for in vitro assay of respiratory supercomplex3-4.

01 January 1970 3,325 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

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

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

Interested to stain mice tissue with WGA 488 conjugate, anybody has a protocol to recommend?

Interested to stain mice brain tissue with WGA 488 conjugate, appreciate if anyone willing to offer me some guidance or suggestions! Thank you!

28 February 2021 3,951 2 View

B-mercaptoethanol (BME) protocol?

Hi everybody, I have been running a multiplex IHC which allow me to stain the same tissue with different markers (Primary Ab + Secondary conjugated Ab) (AF487, AF555, AF647). Since I would like to...

27 February 2021 5,028 2 View

Why do my MCFs not form monolayer?

Hello, I am looking for some expertise on culturing MCF7s. I purchased them from ATCC and I have been following all media and culture conditions. Media is EMEM with 10% FBS and 10ug/mL of...

27 February 2021 3,904 2 View

How can I represent SDS-PAGE results for machine learning analysis?

I am trying to classify and analyze the results of an SDS-PAGE based array for bacterial detection using machine learning, but I have trouble finding the best way to represent the results with...

27 February 2021 9,176 3 View

Is there anyone who experienced with cytotoxic effect of Hoescht 33342 dye?

Is Hoechst toxic to cells? In general, it is known as non-toxic. I am working with dsRED MDA-MB-231 cell line. I applied it for my PDMS-based microfluidic platform, for this reason, I incubated...

26 February 2021 9,119 3 View

Why my agarose gel staining sometimes looks not homogenic but like gradient (see the figure)?

I am worried about this overexposure of the upper part of the gel in the picture. Is it possible that this is the result of too much concentration of ethidium bromide? Why is there such a big...

25 February 2021 8,140 3 View