09 September 2012 2 7K Report

I want to run rAB on gradient SDS-PAGE. I tried twice but the gradient was not formed. Can somebody please send me the protocol?

More Sonalika Mahajan's questions See All
Can somebody please provide suggestions as I am struggling to express ACAP2 protein in mammalian cells?

I have cloned ACAP2 in pCMV-HA vector and the frame has been sequence confirmed. I am trying to express in HEla cells by transfecting he plasmid at 2.5µg per well (6-well plate) using...

08 September 2015 6,482 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 9,296 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 2,858 7 View

Where do I start screening for RNA-protein interactions?

I want to identify the host proteins that interact with the non-coding RNA of virus. Is it possible to do the screening on the similar lines as done in protein-protein interactions using Y2H.

01 February 2015 392 5 View

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

I have estimated the virus titre to be 10 6.5 TCID50/50 µl. I want to know that how much virus I needed to add to infect 25cc flask at 1000 TCID50?

10 November 2014 5,533 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 586 11 View

Can anyone lend advice on CDNA library construction?

I am trying to construct a cell line library for yeast teo hybrid mating. the mRNa concentration was low so I concentrated using a vacuum concentrator and the final conc. of mRNA was 370 ng/µl. I...

06 July 2014 8,294 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 2,238 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 1,422 1 View

Similar questions and discussions
Why I can't see any band in SDS-PAGE?

Currently, when I run SDS-PAGE, I don't see any bands at all, even though I used the same material just a day ago and it worked fine.... In our lab, we dilute the 10X running buffer to 1X and...

06 August 2024 5,373 2 View

Why is my ASO degraded quickly on agarose gel?

I have been working on Red blood cell-derived extracellular vesicles as Antisense Oligonucleotide (ASO) carriers. We normally run agarose gel to quantify the loading efficiency. I used naked ASO...

06 August 2024 3,130 2 View

How to preform densitometry on SDS-page bands?

I ran a SDS-page of a bacterial lysate and I want to quantify protein concentration in a specific band. I was thinking of using a standards ladder or make some standards are different...

05 August 2024 9,805 3 View

I'm optimizing a tetra-ARMS PCR protocol with amplicon sizes: 165bp, 123bp and 93bp. Why, in the gel, only 165bp is visible while I'm expecting all 3?

It's an end-point PCR protocol. I'm using 1.5% agarose gel with SyBR Safe dye and TBE as a running buffer, visualization on BioRad XR+ system. I was primarily thinking of primer efficiency,...

01 August 2024 4,673 4 View

Following click reaction in cell lysates, protein is immobile and remains at the top of the gel in SDS-PAGE?

I am using CuBr/THPTA for a click reaction in total cell lysates. I am facing issues with my protein sample in non-reducing SDS-PAGE where it's not migrating properly and most of it remains at the...

29 July 2024 950 4 View

Why my IgG antibody looks fragmented on non-reducing SDS-PAGE?

I am performing IgG purification and I have to show my results on SDS-PAGE. I use 10% tris glycine gel and prepare the samples under non reducing conditions. I am new to antibodies and therefore...

23 July 2024 6,664 6 View

Why my gel electrophoresis has smearing?

Greetings. I’m currently running a nested pcr for giardia. My mastermix comprised of 3mM Mgcl2, 5unit of taq polymerase, 0.2uM of forward and reverse primers each respectively and 0.2mM of dNtp...

22 July 2024 9,761 5 View

How should we increase the quality of RNA extraction?

I’m having difficulty achieving high RNA integrity in my samples. Although the 260/280 and 260/230 ratios are satisfactory after RNA extraction, the RNA samples show signs of degradation when...

22 July 2024 155 4 View

What is the physical meaning of the magnetic scalar potential?

In cases where the rotational of the magnetic field H is zero, we can define this field as the gradient of a scalar function defined as the magnetic scalar potential (similar to the electric...

21 July 2024 9,633 4 View

What is the cause of this aggregation just bellow 70 kDa mark?

Hello, I was running a 12% SDS Page electrophoresis on few granulosa cell samples and got this result after the ponceau staining. The total protein lysate seem to aggregate at 70 kDa ladder mark...

21 July 2024 5,128 4 View

Our partners will collect data and use cookies for ad personalization and measurement. Learn how we and our ad partner Google, collect and use data. Agree & close