What is the method in detail of reporter gene of luciferase assay ?

I will do the reporter gene by luciferase assay to study the promoter activity?i will use pgl3 basic vector and prl-tk,, the cell line MCF-7and MCF-7 , I will use lipofectamine 2000 and optimum...

04 May 2015 8,038 3 View

Does gene expression of any gene in breast cancer line reflect the gene expression in vivo in the breast ?

when we dealing the cell to convert to cell line , the property of the cell line like gene expression of cell or phenotype from the vivo tissue ? do we can depend the result expression of gene in...

02 March 2015 2,360 9 View

What is the normal level of concentration of TNF-alpha in rat colon tissue ?

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01 February 2015 8,332 0 View

Why did I not got any results with lkb1 primer in real time pcr?

I had mcf-10 cell and was grown in dmem/f12 media with fbs and AB and rna and dna was extracted from the cell ,, when I did RT-PCR with cDNA , I got results with GAPDH but not with my interested...

31 December 2014 9,018 3 View

Why the different size of band with the same gene in pcr ?

31 December 2014 6,189 8 View

What are the reasons for low expression for the gene LKB1 in a mcf-10 cell line?

31 December 2014 8,928 6 View

Can I use pipette tips that have water in them after autoclaving in RNA extraction?

When I autoclaved the tips that will be used in my DNA and RNA extractions, the tips had drops of water inside, although I covered the boxes of tips with foil.

11 December 2014 7,777 13 View

If I isolate DNA and RNA from cell monolayer culture by Trizol, do I need to centrifuge after adding Trizol and before adding choloroform?

11 December 2014 2,515 5 View

What is appropriate media for MCF-7 cell and MCF-10 ?

10 November 2014 1,972 1 View

What is the condition media from cell culture and what are the advantages in using it?

In my experiment, I collected condition media from breast cancer cell culture MDA-231 and I added it to normal breast cell to convert it to cancer, what is the advantage when we used condition...

09 October 2014 8,541 6 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

Buffer to disrupt platelet alpha granules for PF4 quantification?

Hello! I want to quantify by ELISA the secreted (from platelets poor plasma) and the non-secreted (from platelet lysate) PF4 before and after TRAP stimulation. I will use the ELISA from R&D...

03 March 2021 1,499 2 View

Buffer Exchange using column packed with sephadex G-25 effect on protein concentration?

Hello, If i am doing a buffer exchange for an antibody of 1mg/mL, does the elution lose protein in the process of buffer exchange? For example, if i flow through 500 uL of 1mg/mL sample, and...

03 March 2021 6,299 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

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 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

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

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

Anyone that can help with details in phosphopeptides enrichment?

Anyone know the amount of protein before enzymatic digestion for phosphopeptides enrichment of serume sample in Ti-IMAC? Has anyone ever use Ti-IMAC(the material is from Chinese company named...

28 February 2021 7,310 3 View

What is the most suitable resuspension buffer for TCA/acetone precipitated protein?

I need to extract protein from fermented carbon sources for Bradford assay. Most researchers experiencing insolubility of pellet in resuspension buffer. Please assist me to select most suitable...

26 February 2021 7,129 3 View