How much spread is acceptable in relative gene expression in qPCR experiments?

Hi, I have recently done some qPCR after total RNA extraction, followed by first strand cDNA synthesis. The problem is, while one treatment is giving me closely lying values for relative gene...

04 May 2015 722 6 View

How can I study DNA-protein complex with AFM?

Hi, I want to study the structure of the DNA-protein complex using AFM. The problem is I don't have purified protein with me. So, as an alternative, I am planning to incubate the DNA fragment with...

01 February 2015 5,188 3 View

How can I predict DNA secondary structure?

I have a DNA sequence and I suspect that it forms a curved structure, however, while I am planning the experiments to test this hypothesis, I would like to do some in-silico prediction to help me...

01 February 2015 1,539 3 View

Can 1% TritonX-100 be used instead of 0.1%Nonidet-P40 for Co-IP lysis buffer?

Hi, I want to know if TritonX-100 can be used instead of 0.1%Nonidet-P40 7 for Co-IP lysis buffer? The proteins I am targeting are both nuclear proteins.

10 November 2014 1,388 9 View

What is the best kit for total RNA extraction from yeast?

HI, I am using the TRIzol reagent for RNA extraction from yeast. I am planning to shift to a kit based protocol for better reproducibility and consistency. I looked for many RNA extraction kits...

10 November 2014 539 8 View

What reference genes are best for qPCR with S. cerevisiae cDNA smples.

Hi, I am doing qPCR assays with S. cerevisiae. As per the article "Validation of reference genes for quantitative expression analysis by real-time RT-PCR in Saccharomyces...

10 November 2014 5,825 11 View

anti GBD and anti GAD antibodies?

Hi, I have to see some protein-protein interaction for which I am using the yeast two hybrid system. After constructing the fusion GBD and GAD constructs (in pGBD C1 and pGAD C1 vectors) I want to...

11 December 2012 1,433 0 View

Why does blocking with my immunogenic peptide cause nuclear fluorescence in ICC?

Hi all, I am validating a polyclonal antibody raised against human ANT4, through WB and ICC. In most of my controls, the antibody shows very little off-target binding, but when I pre-incubate the...

03 March 2021 5,999 3 View

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02 March 2021 3,060 3 View

Does Napabucasin soluble in dichloromethane and in ethanol?

I need to prepare a solution of Napabucasin (inhibitor of cancer cell stemness) in dichloromethane. The drug is expensive, so maybe somebody already tried to do it. If it is soluble, what is the...

02 March 2021 9,945 3 View

Help request in CST microwave?

im working now on TPV solar cells and metamaterials and i realy need our help in CST microwave i have a tpv cell simulation and i want to extract particullarly the absorbtion of only the active...

02 March 2021 2,246 2 View

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

What do I do with DLS peaks that are from the buffer?

I'm looking at the aggregation of my protein sample using DLS. Unfortunately, my buffer (20mM HEPES) also results in a set of peaks. These are at approximately 1 and 1000 d.nm. The lowest peak...

01 March 2021 9,015 2 View

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View