Drawbacks of Gene Reporter assay?

More Ahmed Khalaf's questions See All

Should we normalize sum of two drugs' effect when i compare it to cobination effect in cell lines?

i am analysing data obtained from technical experiment of treated same count of cell lines with two different drugs and their combination. Then i need to find out whether there is a synergy for...

02 March 2019 8,700 4 View

What is the approved method to induce EGFR T790M in mice chemically ?

What is the approved method to induce EGFR T790M in mice chemically ? I am trying to find toxic chemical or drug that has the ability to induce carcinogenicity through double EGFR mutation (L858R...

31 December 2017 435 1 View

Where can we get SUMO-1 Plasmid?

Could our researchers, working on a plasmid SUMO-1, supply us with it?Thanks in advance

11 December 2015 7,985 2 View

What is the best method to make IP for HA tagged proteins?

Could you provide me with a protocol for best method of HA pulldown?

11 December 2015 5,803 5 View

What is the order of COD kinetics?

our cell line will be transfected with 2 different plasmids at the same time for 48 hours but the results is not equal between all wells and the results should show us an equality so my question...

11 December 2015 3,920 3 View

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 View

RIA. How to measure?

When you use RIA, with a control the unknown sample with antibodies, you bare in mind the binding sites of the antibodies. However you still need to measure labelled antigen (radioactive) and not...

28 February 2021 2,133 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

EMSA troubleshooting. Protein-DNA complex not migrating, cold probe lane with multiple bands. Can anyone help?

I'm running an EMSA to show the DNA binding activity of recombinant proteins versus the Wild type. Previous assays run by my research group using a different test system show the protein-DNA...

24 February 2021 8,325 1 View

How we can measure Kemp % in wool without using of benzene?

Would you please let me to know about the methods for measurement of medullation and Kemp % on animal? My problem is that the wool sampling and sending to laboratory is not possible. So I would...

21 February 2021 7,260 4 View

What is the proper in vitro culturing method to get information about the activation of innate immune cells by VLPs?

Dear All, I am interested in studying the response of immune cells against virus-like particles (VLPs). I used the PBMC in vitro assay and treat it with the VLP using PHA as a positive ctrl while...

16 February 2021 1,337 2 View

Why does flow deformation occur at the test line and control line on nitrocellulose membranes (LFIA)?

I use 1% BSA in 10 mM phosphate buffer (pH 7.4) solution when striping my nitrocellulose membranes with antigens and antibodies. However, I always observe deformed flow when my sample or running...

16 February 2021 4,224 3 View

Can the viability assays (ex: MTT/CCK-8) measure cell proliferation? or are they solely viability assays?

I want to conduct a proliferation assay to test a chemotherapy combination on a cancer cell line. At first, I thought about the MTT assay. But now, all sources are saying that these do not measure...

14 February 2021 3,142 7 View

GST assay in microplate?

I'd like to know if anyone can provide me with a protocol for traditional GST enzymatic assay (with GSH and CDNB as substrates) in a 96 well microplate, for cell line and/or C.elegans...

11 February 2021 1,328 3 View

What lysis buffer is compatible with Bradford assay?

Hello! I am planning to use Bradford assay to measure protein content in PBMCs lysates. The problem is that Bradford assay doesn't seem to be compatible with Tris-based buffers or SDS. Are there...

10 February 2021 1,797 5 View

Reiner Andag

I think it is better to use Dual Luciferase Assay from Promega.If you are using this method you can measur Firefly- and Renilla -Luciferase in one tube.

Ahmed Khalaf

Thank you, I already underway of using Promega but without dealing with Renilla, B-Gal instead.

But what is technical drawbacks ?

That you have two different Assays.Using dual Luc,1.measur F-Luc ,then stop reaction and add substrate for R-Ruc.The advantage is,that you can`t make mistakes in dependence of the sample volume.