How do I generate a generic protein motif?

Hi everyone, I am currently working on a project that requires me to generate a generic motif that I can blast against my database of interest. Could someone guide me on how this can be done?...

31 December 2014 9,028 3 View

How are conditions for bacterial cell sonication determined?

Hi guys, I am currently optimising the expression of a protein that is found predominantly in the pellet. However, I am unsure how the sonication condition can be adjusted such that I have a...

11 December 2014 2,018 8 View

A query on protein purification and refolding from insoluble fraction

Hi, I recently purified an eukaryotic protein from BL21 cells. Upon examining the fractions, it seems that this protein is found predominantly in the insoluble pool. As such, I need some advice on...

09 October 2014 7,184 8 View

What is the purpose of Tetrabutylammonium bromide in HPLC?

I chanced upon a HPLC protocol that uses acetonitrile, Potassium phosphate and tetrabutylammonium bromide as the solvent for reverse phase chromatography. I believe potassium phosphate is most...

08 September 2014 4,221 14 View

Can IPTG induced cells be kept in the pellet form at minus 20 degree Celsius for prolonged period before cell lysis?

I heard that once the supernatant of cell culture is removed, the pellet can be kept in minus 20 fridge. However, I wonder if this can be done for long period of time. Also, please suggest if...

08 September 2014 7,356 7 View

What are the recommended ways to preserve proteins that are to be subjected to downstream activity tests?

Hi everyone, may i know how proteins can be preserved upon purification? I need these proteins for downstream activity tests but as I will not need them right away, I need some solutions to...

08 September 2014 8,384 7 View

What considerations should I make over the types of salt present in buffers?

Hello guys, may I know what are the considerations one should have when it comes to the selection of buffers for protein purification? I learnt that optimal pH range and components such as...

07 August 2014 5,430 5 View

Any advice on reduced yield of purified protein upon dialysis (Pierce slide A lyzer) and concentrating (Amicon 10K)?

Hello guys, I have recently purified a his-tagged protein via Ni beads. Based on a BSA standard, it was estimated that the eluted protein concentration was about 1 microgram per microlitre in 100...

07 August 2014 2,141 10 View

Is the loading buffer (lamelli) sufficient for removing proteins that are pulled down by Nickel beads for western blot?

07 August 2014 4,427 3 View

How can I estimate a protein isoelectric point and its implication on purification?

Hi guys, I would like to know if there is any recommended websites that I can rely on for isoelectric point estimation? I used the one by expasy and it seems that the value deviates by at least...

06 July 2014 3,038 20 View

Adherance of Polyaniline coating over stainless steel?

Is the electropolymerisation of polyaniline coating over stainless steel (SS) via Cyclic voltammetry technique is really adherant? Else how to improve the adhesion of the coating, whether any...

01 March 2021 8,220 2 View

Is there a method of determining feed digestibility without the need to re-pellet the feed?

I am aiming to determine the ADC values of a commercial fish feed, but I do not have a steam pelleter or extruder available to me. Ideally, the pellets could be coated with some inert marker....

28 February 2021 7,967 3 View

How to immerse bricks in wax ( for coating ) ?

Do we immerse the bricks horizontally or vertically ? #bricks#waterproofingsurface#civil#engeenering#chemicalengineering#wax#parafinwax

28 February 2021 6,685 2 View

For gelatin coating for culturing HUVEC.Should the gelatin be made in sterile water or PBS? Do I need to autoclave the gelatin soln after making it ?

I want to use 0.1% gelatin. Should that be made in PBS or sterile water? Should I autoclave the gelatin soln after making it in water/PBS?

28 February 2021 274 2 View

Why is my platinum counter electrode getting dirty during electrodeposition?

I am conducting anodic electrodeposition of MnO2 onto a stainless steel substrate, using a platinum strip as a counter electrode. This is done galvanostatically, with the current held between 5...

27 February 2021 1,829 3 View

What pH condition required to coat APTES on silica coated Fe3O4?

It's a hydrolysis reaction, which may produce by either acid or base-catalyzed reaction. so can anybody help me with this, which condition is suitable for this reaction?

26 February 2021 1,760 8 View

What is the best surface to culture cells isolated from a skin biopsy?

Hello, recently I am having some difficulties with maintaining the cell culture of primary cells isolated from a skin biopsy (normal and malignant). The coating of culture plates with Matrigel (a...

26 February 2021 6,151 2 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

How does charge on working electrode changes with rising potential?

During performing CV of a GCE or modified GCE, we rise the potential in the first half of the cycle and reduce it in reverse half. During the first half, what is the nature of charge on GCE or...

25 February 2021 7,994 3 View

What condition required for coating of Fe3O4@SiO2 by using APTES?

I tried coating Fe3O4@ SiO2 by using APTES in 5% water-ethanol but it is not coated. Kindly suggest a solution for it. Thanks

24 February 2021 5,992 4 View