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...

31 December 2016 2,402 4 View

Indexing and DOI for an article?

06 July 2016 4,791 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...

11 December 2014 3,277 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%...

11 December 2013 4,850 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...

03 April 2013 6,006 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...

02 March 2013 2,718 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...

31 December 2012 167 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...

09 October 2012 8,252 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...

08 September 2012 7,705 35 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...

07 August 2012 5,373 14 View

MiRNA qpcr problem?

Dear All, mirna primer showing some problem in the melting curve? any idea why? As attached is the melting curve. The forward sequence is obtained from miRBase and reverse primer is universal.

28 February 2021 5,008 4 View

Can MicroRNA cross the blood brain barrier?

In diseased conditions, many genes and gene products are visible in the bloodstream. So my question is whether we can find traces of microRNA which is involved in disease and disorder in blood serum?

24 February 2021 7,064 3 View

Are exogenous molecules transferred between human cells during doubling ?

Hello RG community, I wonderer whether exogenous molecules (e.g. reporter plasmids, miRNAs or other artificial molecules) are transferred between cultured human cells during their doubling ? So,...

21 February 2021 6,633 3 View

Has anyone encountered any problems with abm good microRNA primers for quantification of mature miRNAs?

I've been trying to quantitate mature microRNAs in 293T cells with and without 'X'. 2ug of total RNA (isolated by Trizol method) is converted to cDNA using All-in one microRNA cDNA synthesis kit...

17 February 2021 8,986 2 View

What is the best way to fix plasma derived miRNA for subsequent cDNA synthesis?

I need to quantify blood plasma derived miRNA through q-RT-PCR. For this I need to fix RNA content in my cDNA reaction. What is the best way to do this? Currently, I take readings of my plasma...

03 February 2021 9,330 3 View

What is the general miRNA yield one gets from the Qiagen miRNEASY serum/plasma advanced kit?

I am using the Qiagen miRNEASY serum/plasma advanced kit, using 400ul of frozen plasma. What is the yield you get on average? I am using nanodrop currently for quantification. I am getting...

02 February 2021 8,806 2 View

Which miRNA arm is a right target for analysis, 3p or 5p?

I'm working on selecting mouse miRNAs to analyze and stumbling upon one confusing issue, some of the papers do not specify which arm they analyzed and they just state something like " miR-34a was...

16 January 2021 3,792 4 View

Is negative controls necessary for western blotting?

I am trying to confirm my miRNA target by western blotting. I have a ds mimic and a ss inhibitor of my miRNA. Is it necessary to use negative controls for mimic and inhibitor? If yes, how can I...

09 January 2021 2,320 3 View

Can I predict miRNA by knowing the mRNA target?

Hello! I am trying to predict in silico the miRNA by knowing the target. Most of published research starts by knowing the miRNA and predicts the target using online tools. Are you familiar with...

31 December 2020 7,433 5 View

What are the possible drawbacks of using miRNA sponge to enhance the specificity of gene expression?

I am searching for miRNA repeats put between WPRE and 3'LTR of my lentivirus, in order to enhance the specificity of transgene expression in human neuron cells. But if clinically translation of...

26 December 2020 9,921 2 View