Shahid Mahmood

11 Questions 18 Answers 0 Followers

Questions related from Shahid Mahmood

How to get better resolution of protein with Ion exchange chromatography?

Protein Profile: Recombinant protein subunit expressed in bacteria. Theoretical pI: 8.0. adaptor protein. IEX Profile: Anion exchanger (Hitrap Q FF Column 1ml) Running conditions: Start buffer:...

25 February 2025 8,425 5 View

How to chemically enhance the Ecoli BL21(DE3) membrane permeability against negatively charged unnatural amino acid (sTyr)?

hi, I am expressing protein with Unnatural Amino acid (UAAs) especially negatively charged UAAs. i am trying different -ve UAAs they were easy to incorporate in protein and gave good expression....

26 September 2024 8,070 5 View

Have different band with same protein after doing Ion exchange (IEX)?

I expressed UAAs incorporated protein. for sake of getting pure protein i run IEX. as shown in picture from left to right LANE#1, LANE#3, LANE#4 is same protein and the concentration is 1.6mg/ml,...

20 June 2024 4,648 4 View

How to prevent recombinant protein from GSH abduct incorporation?

Hi, experts. I'm trying to expressed recombinant transmembrane protein incorporated with nucleophilic non-canonical amino acids(ncAAs) by genetic code expansion (GCE) in E.coli. I replace tyrosine...

04 February 2024 2,500 0 View

How to inhibit Glutathion(GSH) protein modification in E.coli?

Hi, i am working on unnatural Amino Acid (UAAs) incorporated Insulin receptor substrate (IRS-1) expression and purification in BL21(DE3) strain. While i expressed my protein with UAA Glutathion...

24 January 2024 1,484 4 View

How resolve if i got WT peak in Mass results?

Hey Researchers, i am working with transformation of membrane protein by using BL21(DE) competent cell. i transformed plasmid (pET28a(FLAG, His tag) and co-transformed with pULTRA-ambryx. i...

27 November 2023 3,618 0 View

How do i optimize the Phage-antibody competitive ELISA?

Hello, I am working on scFV phage display library. I got positive monoclonal from Phage display library. Now i did Phage-antibody competitive ELISA. i got following surprising results. 1) Phage...

30 October 2023 205 0 View

Got short fragments in Colony PCR of Phage display lib panning random pick?

I'm doing scFv phage display library panning with photocross linked protien. after panning i performed colony PCR for confirmation of scFv length. i got various short length fragment on gel, only...

06 July 2023 8,160 2 View

Why Chemiluminescence Substrate (ECL) left color with 1 minutes after adding on WB membrane?

I did Western Blot (WB) with Photo-cross linked protein. I had some unusual thing (maybe for me). When i added ECL substrate on membrane. The signals were getting weak with in just 1 mint and...

01 June 2023 7,037 3 View

Recommendation for miRNA expression analysis for Type 2 Diabetes?

Which techniques are more accurate and far-better for miRNA expression analysis. Is micro array or miRNA sequences tech? if RNA Sequence tech then please share article for initial understanding or...

12 March 2018 2,812 3 View

What is the alternative of Pen-Strep in cell culture?

Anything or alternative antibiotic that i use instead of Pen-Strep. Any other antibiotic that use for cell culture.

27 October 2014 9,939 5 View