Ahmed E. Dhamad

9 Questions 85 Answers 0 Followers

Questions related from Ahmed E. Dhamad

What does it mean when you have less cells# in flow cytometry plot compared to a control group?

Hi! I have run flow cytometry for 3 different cell lines (black, blue, and red) which were stained with red dye (FL2) with a same event number (15,000 events). However, the cell number (count; Y...

17 October 2017 2,995 3 View

Is there any difference regard to a cell behavior after knock out a gene using different plasmids ?

CRISPR-Cas9 has been used to knock out a gene in breast cancer cell (MCF7) by using two different plasmids. One plasmid has a puromycin selection marker and another one has a GFP selection marker....

01 February 2017 2,340 2 View

Is there any ideas why the over expressed protein behaves differently than the endogenous protein?

I over-expressed HA tagged mammalian protein (27KD) in 3 different cell lines; however, I saw the over expressed protein (exogenous) behaves differently than the endogenous protein in the all cell...

11 October 2016 7,497 8 View

What is the best way to test several genes expressions using qPCR?

I would like to determine several gene expressions for 4 different samples using qPCR with SYBR Green. Is it possible to run all of them in one plate regard to primers Tm differences? Would run...

10 July 2016 4,271 3 View

Which are the amino-sequence cutting sites of bovine thrombin?

Hello, I am trying to cleavage my protein from GST-tag by using Thrombin with its site ( L-V-P-R-G-S) which is provided by plasmid. However, I am confused because I saw after running my protein...

10 November 2014 8,152 9 View

Can I separate two proteins which have almost the same M.Wt. and isoelectric points of 7.1 and 7.75 using IEC?

I have two proteins which have almost the same M.Wt. They also have isoelectric points (7.1 and 7.75). Can I separate them with Ion exchange chromatography(IEC)? What is the PI range by which Ion...

08 October 2014 1,071 6 View

Does anyone perform S-Carboxymethylation of Cysteine for protein or peptide? I am looking for the protocol . Any help would be appreciated

I am looking for the S-Carboxymethylation of Cysteine protocol. I want to methylate my protein by using iodoacetic acid. If you have a procedure for that, please send my a copy or tell me where I...

19 September 2014 7,907 5 View

Is it important to know how many passages of the cell line have passed?

I am working with MCF7 and 293T cells. Do I have to know the # of passages that they have passed? If yes, what is the maximum passage's number, in general to my cells?

21 May 2014 3,061 17 View

How can I get rid of a non-specific binding with GST- beads?

I am doing protein mapping to my interesting protein (chop it off for different domains, mix them with other GST- protein to do in vitro binding assay and then they are pulled down with GST-...

12 May 2014 9,836 7 View