Mohamed Alalem

14 Questions 21 Answers 0 Followers

Questions related from Mohamed Alalem

Any Idea please as where to find customized Ab that does not bind to protein A?

I need customized Ab against some antigens and I would like to have isotypes/host variants of the Ab that does not bind to protein A. Is there a company that I could call to discuss customization...

01 January 2017 2,425 4 View

Indexing and DOI for an article?

How to assign a DOI for an article, please, does anyone have an idea as how to obtain a DOI for my article. Thank you Best regards

07 July 2016 4,817 5 View

How could one guarantee in digital PCR that always some partitioned droplets will have no template and will be zero?

Digital PCR relies on proportion of positive and negative results in the droplets. What is the guarantee that not all droplets will have atleast one template and thence all droplets will be...

12 December 2014 3,299 4 View

Any advice on Fluorescent EMSA?

These dCTP fluorescent nucleotides were incorporated into a DNA probe and I ran the free probe in the first lane from the left and cell lysate with the labelled probe in the second lane in 6%...

12 December 2013 4,877 3 View

Loading equal cell extract volume instead of equal amount of protein

I would like to know what is the difference and, the advantages and disadvantages of loading equal volume of cell extract instead of loading an equal amount of protein per lane in SDS-PAGE. I am...

04 April 2013 6,031 10 View

Phenol Extraction of DNA.

I have done phenol extraction and ethanol precipitation of a DNA sample. I have run the DNA on agarose 1% gel and excised the desired band. The band showed good intensity. The excised band was...

03 March 2013 2,739 15 View

Finding the introns as well as the 5' and 3' UTR for a given gene from the cDNA

I would like to find the genomic sequence of a given gene. I have used the NCBI Blast for a sequence from the CDS of the gene and it gave hits corresponding to the location of this sequence on...

01 January 2013 188 9 View

Quantification of protein band density in X-ray films from ECL Western Blots

I have used Image J, Photoshop and Vision Works software to compare protein band densities in x-ray images from ECL-WB, yet there was a huge discrepancy among the results. I think that...

10 October 2012 8,278 18 View

ELISA for a protein secreted by culture cells into growth medium

Does anybody know how to use an ELISA to test for proteins secreted into DMEM growth medium by culture cells? I have the antibodies to these proteins and I would like to know whether I should...

09 September 2012 7,725 35 View

Lysis of cell membrane in cell debris using RIPA lysis buffer to extract membrane-bound proteins from the cell debris

I am planning to use RIPA lysis buffer on cell debris to study membrane-bound protein fraction. Yet, I found that mere vortex of the cell debris in RIPA buffer is not sufficient to release...

08 August 2012 3,096 4 View

Nonspecific black dots on PVDF after ECL in WB.

Please see attached image of the X-ray film after ECL of the PVDF in WB. The Arrows indicate the black non specific spots in the lower part of the image and the non specific bands which do not...

08 August 2012 5,401 14 View

Solubilization of eukaryotic cell membranes from pelleted cells

I would like to load a membrane fraction of eukaryotic cells into SDS-PAGE gel. What is the best buffer I can use to solubilize the eukaryotic pelleted cells?

07 July 2012 1,421 2 View

Nonspecific black dots on PVDF after ECL in WB.

I have stripped a PVDF and blocked it in 5% nonfat milk and reprobed it with another primary Ab and proceeded with the washing and secondary Ab incubation, but I had nonspecific black dots on the...

06 June 2012 4,998 26 View

Transfection of mammalian breast cell lines by miRNA and shRNA to stably silence a given gene.

Hi, I would like to know, please, what is the best plasmid vector design I should use to stably express miRNA or shRNA in mammalian breast cells. Thank You!

01 January 2012 881 3 View