Hello, my protein is undergoing aggregation and eluting in void volume during gel filtration. How can I reduce this ?

After affinity through amylose and TEV cleavage, the protein is aggregated in GPC so I am not getting any active protein out of this. How can I minimise aggregation.

03 April 2018 1,046 0 View

Hello < I am trying to standardize IFA with Plasmodium falciparum parasitized RBCs . Can you help in that?

And am using 1:400 dilution for my protein's antibody. With overnight 4 deg binding. Should I make it 1 hour at RT. I increased tween in PBS from 0.01 to 0.05 % . I got some less background. But...

09 October 2017 1,251 0 View

Can you give me a protocol for GST assay?

I am doing it with the sigma GST assay kit. My blank buffer consisting of PBS, GSH and cDNB is also giving an absorbance. So can you just give me a detailed description in case am making a mistake.

07 August 2017 8,894 2 View

How to get rid of chaperone at around 63 kD during recombinant protein expression in C43 competent cells?

I was inducing with 0.8 mM IPTG and kept 16-18 hrs post induction at 25 degree..am trying to reduce the temp and check the expression ; also lowering the salt and increasing imidazole...

06 July 2017 8,807 2 View

Does HIC help to get rid of chaperones?

06 July 2017 8,799 5 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

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

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 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

What urea buffer can i use for biotagged protein extraction from streptavidin beads?

Hi all, I have been doing research on biotagged LDB1 proteins in Mel cells. I purify the proteins using M280 streptavidin beads. The yield is very low so I've been trying to optimize it by...

24 February 2021 5,749 3 View

Quantifying leaking DNA from bacteria - what's the secret?

Hi, all! I need to determine if my treatment causes membrane damage in bacteria (S. aureus and E. cloacae), but I cannot use a fluorescent probe such as PI and similar ones because my compound is...

24 February 2021 2,006 2 View

Is there an ideal incubation time of RGO-modified SPCE before subsequent use, i.e., electrochemical characterization or further modification?

I notice that upon washing and air-drying of my RGO/SPCE surface and characterizing again via DPV, the current response towards ferricyanide probe significantly decreases. My worry is that the...

22 February 2021 5,324 3 View