Fatin Safiudin

12 Questions 8 Answers 0 Followers

Questions related from Fatin Safiudin

How to fix partially purified protein precipitation issue?

I keep all my partially purified protein in the respective elution buffer (20mM NaP, 0.5M salt, 100mM imidazole) at -20° freezer. I plan to do a buffer change today for my next polishing step....

07 July 2023 4,940 2 View

Do I need to take into account the protein tag when calculating isoelectric point (pI) of protein for ion exchange chromatography?

Should I take into account the protein tag (His/ GST) during the calculation of pI to choose which ion exchanger I should use? Or should I consider only the real target protein sequence only?

05 July 2023 1,458 2 View

Does my GST protein tag hidden in the structure?

My N -terminal GST tagged protein fail to purify using this condition: 30ml sample (3mg of crude protein) injected at flowrate (0.5ml/min), 40ml wash (PBS pH 7.3) and 5ml elution (50mM Tris HCl +...

06 June 2023 5,697 4 View

Is my protein stuck in the well during Native -PAGE?

I performed zymogram analysis for lipase using this protocol: 1. Run Native -PAGE until adequate migration (roughly 80V for top gel: 30min, 120V for bottom gel: 2 hour) at 4 °C. Cathode connects...

06 June 2023 6,598 5 View

Why does my pNPP substrate changed color?

My study is on lipase and I planned to used p-NP palmitate method as the activity assay. The recipe includes Tris HCl buffer pH 9 + Triton X-100 + isopropanol. I stirred the mixture with heat then...

22 March 2023 3,268 4 View

What is the best/ most convenient/ cheapest alternative for an automated purification system like Akta purifier?

I was looking for any recommendation for protein purification system for my research. We dont have much budget for akta and are looking for affordable alternative. We do have HPLC in the lab, can...

08 March 2023 7,555 3 View

Does my protein bind to the resin?

I conducted gravitational His tag protein purification manually and observed that the A280 of my crude is similar to the eluent post sample application. How is this possible? Does the His protein...

30 November 2022 1,619 5 View

Can I reuse 50ml DURAN glass bottle after storage of 0.1M NiSO4?

Ive made a mistake of preparing 0.1M NiSO4 in DURAN glass bottle. Can I reuse the bottle? If so, how can I neutralize the bottle from any traces of NiSO4?

17 October 2022 1,857 2 View

How many Cys residues for my protein to be considered as cysteine rich?

Hi, I couldnt find anywhere the exact percentage or amount of cysteine need to be present for my protein to be considered as cysteine rich. May I know the exact number/ percentage? for example >1%

25 August 2022 8,245 2 View

Why do I get different result for the prediction of protein secondary structure?

I did an in silico analysis for my protein to understand its secondary structure by using two servers; PSIPRED Workbench (http://bioinf.cs.ucl.ac.uk/psipred/) and SOPMA...

22 August 2022 409 3 View

Why do my protein fail to express?

Hi, recently I tried to express my genes (from plant) into Rosetta De3 pLysS using this protocol: 1. 1% overnight culture into a new media + antibiotic, incubated to grow until OD600= 0.5 2....

11 August 2022 2,573 1 View

Can I reuse my SDS gel?

My gel set can hold 4 gels but my samples can fit in 2. I have only one dummy plate so I have one extra empty slot. Can I use an empty SDS gel as dummy? And can I reuse the gel back with sample?

02 June 2022 1,826 2 View