Can somebody please provide suggestions as I am struggling to express ACAP2 protein in mammalian cells?

08 September 2015 6,386 3 View

How can I express GST-tagged recombinant protein in soluble form and prevent the inclusion body formation?

I have expressed GST-tagged protein in BL21(DE3)pLysS cells but I am not able to get the protein in soluble form. Most of the protein is in inclusion body. The induction conditions used are 1mM...

04 May 2015 9,202 9 View

What is the best method to lyse bovine kidney cell line?

I want to perform Co-IP from the bovine kidney cell line transfected with the Myc-tagged bait and HA-tagged prey proteins. I am using Pierce RIPA lysis buffer for the cell lysis but the lysate I'm...

02 March 2015 2,717 7 View

Where do I start screening for RNA-protein interactions?

01 February 2015 296 5 View

How to calcuate the dose of virus needed to infect 25cc flask at 1000 TCID50?

10 November 2014 5,412 3 View

Can somebody please suggest some tools to perform Gene ontology t0 the third level using UniProt IDs of the genes ?

I have screened a cDNA library of swine. I want to perform gene ontology on the obtained gene sequences so that a pie diagram representinbg the proportion of genes participating in various...

09 October 2014 475 11 View

Can anyone lend advice on CDNA library construction?

06 July 2014 8,182 4 View

Can somebody suggest any alternate method of Acetone protein concentration and analysis?

I have expressed one viral gene in Pischia pastoris. the expressed recombinant protein is secreted in the supernatant. I have concentratexd the supernatant using acetone and TCA method and run the...

03 April 2014 2,121 3 View

Has anyone expressed Nus tagged fusion protein?

I am trying to express one protein in pET-43.1 vector fused with N-terminus Nus tag but the size of the expressed protein is about 20 kDa smaller than expected. I also transformed the neat...

03 April 2014 1,318 1 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

Gel electrophoresis result for total RNA samples, 1.5 Agarose gel was used, How to improve the integrity of RNA?

Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue

02 March 2021 5,433 5 View

How to regulate SSCP temperature in simple PAGE?

I'd like to perform single-strand conformation polymorphism (SSCP) in my thesis, however I cannot control the temperature of the vertical PAGE since we are using the conventional tanks. Is there a...

02 March 2021 9,157 1 View

Why are my bands weird in this Agarose gel?

Can someone please give me some possible things that could go wrong? Here is my recipe: 0.5g Agarose 50 mL of TAE 1x 1 uL ethyl bromide. Gel was run at 100V for 1 hour. The buffer used is also TAE.

01 March 2021 9,952 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 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