Why I amGetting non specific bands for my MBP fusion protein after GPC?

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Double digestion problem ?

In order to do a cloning I am trying to do double digestion of my vector with Sph1 and Not1 enzymes , the sites are 14 bases apart. My question is - is it appropriate to do double digestion at a...

03 April 2019 562 3 View

How much concentration of template(Rec Plasmid) required to see a fallout of 166bp?

Hi all, I m very much confused for a double digestion reaction to check a fallout of 166 bp which has been cloned in 6900bp vector. I have took around 1500 ng of my recombinant plasmid(6900+166...

03 April 2019 6,999 4 View

How can I get Chromosomal location of fungal genes?

Hello all, I am looking for bioinformatic tool/websites to know the location of fungal genes, as most of the genes are uncharacterized , is it possible to get their location on chromosome? My...

02 March 2019 3,372 2 View

Any suggestion why i am getting higher protein band size??

Hello all! I am doing SDS-PAGE for my protein of predicted size of 53.5kda bt unexpectedly I am getting bands of around 60kda.. I have done this 3-4 times and Everytime the required band is about...

01 February 2019 5,566 8 View

Does Mutation In Ribosomal Binding Site cause any interference in protein Expression?

Does single nucleotide mutation in Ribosomal Binding Site of an expression vector hamper protein expression? As there is no band could be seen in SDS-PAGE..

07 August 2018 4,068 5 View

My clone ( from cDNA) after sequencing reveals 2bp deletion in , should I use SDM for repair?

My clone ( cDNA Clone) has 2 bp (C&A) deletion as a result their is frame shift and no protein expression is taking place. gDNA clone reveals no deletion but all my cDNA clone reveals 2bp...

06 July 2018 10,000 9 View

I m trying to clone one gene from cDNA(502bp) and gDNA(600bp), during PCR i m getting amplification from gDNA only not from cDNA, any solution??

Same cDNA is used for other amplifications and i m getting result for that ,, actually the Tm of the primers is too low unfortunately it is around 35 degree , when i put an PCR at Tm 28 i got some...

01 February 2018 6,688 6 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

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

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

Are protein samples in SDS compatible with ELISA?

I have used an AllPrep DNA/RNA/Protein Mini Kit QIAGEN kit to extract RNA and protein from my samples. At the end of the protein purification, I resuspend my protein in 5% SDS. Will these samples...

28 February 2021 7,370 3 View

For modelling epoxy-based matrices (in composites), which model in ABAQUS is the most suitable, Concrete damaged plasticity or Drucker-Prager model?

I am working on developing a micromechanical FE model for predicting kink-band formation in UD composites. To model matrix plasticity, which model, out of Concrete Damaged Plasticity and...

28 February 2021 5,415 1 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

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

Why tick's salivary gland extract didn't run in the gel (SDS PAGE )??

Greetings, I want to make SDS PAGE for my sample (Tick salivery gland extract), I tried concentrations 10%, 12%. I also tried high molecular weight ladder. But the protein stopped at the top and...

25 February 2021 7,698 3 View

Jasim Al-saadi

I think you have proteolysis and breakdown of your protein into smaller polypeptides or amino acids . try to inhibit proteases before purification.

Shubhashish Chakraborty

Their is always a problem with MBP. I would suggest you if possible use some other tag such as His or GST. Mostly even after cleaving MBP the protein remains bound due to high affinity of MBP. Good luck

Ravi Shankar Gautam

Sebastian Schmitt

i have N-terminal MPB tag and had used Protease Inhibitor and it does form dimer and oligomer.