miRNAs used: mimics for both 125b and let7B (will work with inhibitors also since this is a product of an endometriosis paper done previously)
transfection reagent: RNAimax
If you want to detect the alterations in the cytokine abundance I would suggest to perform an Elisa or Elisplot with your treated macrophages. This would be the best way to see if your treatment has a functional consequence on them.
03 April 2016 8,370 3 View
Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...
01 March 2021 3,867 3 View
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
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
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
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
Hello, I am an undergraduate student doing my dissertation, where I isolate PBMCs from leucocyte cones in order to plate monocytes and allow for macrophage differentiation. Although the numbers...
09 February 2021 5,334 8 View
Hello, I'm trying to identify good putative macrophage markers for flow cytometry in rats.
04 February 2021 3,913 4 View
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
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
I culture RAW 264.7 cells on my samples and differentiate them to osteoclasts with RANKL. I want to do resorption pit assay by culturing cells for 28 days. My problem is that the macrophages keep...
01 February 2021 6,746 2 View