In situ hybridization on cre-recombinased mice protocol?

Has anyone done FISH on cre-recombinased mice? I want to know if anyone has successfully been able to quench the autofluorescence from the those mice to successfully visualize the targeted genes?...

31 December 2019 5,028 3 View

Liquid chromatography mass spectrometry (LC-MS) and condition media analysis?

Has anyone done LCMS on condition media for protein quantification? I have this media (after the exposure of different flow conditions) that i would like to use LCMS for protein quantification...

01 February 2018 1,344 8 View

Porcine vascular endothelial cell isolation - from the aorta?

Hello, Are there any protocols for the isolation and purification of porcine vascular endothelial cells (from the aorta)? I would like to have my study from the primary culture instead of...

09 October 2017 9,578 3 View

Condition media assays?

Hello, I was wondering if anyone knows different conditioned media assays to quantify protein concentrations besides using ELISA. I have conditioned media from arias treatments and I want to do...

08 September 2017 3,674 2 View

RT-qPCR data analysis..(deltadeltaCt method)?

Hello, I have three different groups (Static/A/B) and I want to use delta delta Ct method to show my results in a log form. Previously I always had an untreated control to do the delta delta...

01 February 2017 7,676 3 View

Any good ALPL primer set for porcine?

01 February 2017 386 1 View

How to perform cDNA and primer quality assessments?

Hi, I have recently been trying to run RT-qPCR experiments for interstitial cells. I used Qiagen RNA extraction kit where I got ~500ng/ul of RNA with 260/280~2 and 260/230~1.9, which shows me that...

31 December 2016 7,986 12 View

How to avoid interstitial cell contamination from endothelial cells?

Does anyone know of a method/tip to avoid interstitial cell contamination from endothelial cells? I have been trying to isolate valvular endothelial cells (pig) and keep getting interstitial cell...

11 December 2016 8,311 4 View

Where to start learning molecular biology/signaling pathways?

I want to start learning about different signaling pathways and post transcriptional pathways.. I am not sure where to start. Any help/tip/resources would be greatly appreciated

11 December 2016 4,716 2 View

Any software recommendations for digitizing images?

Does anyone has any software (free) recommendation for digitizing images? I have few echocardiogram images (velocity vs. time) that I would like to digitize. I have been trying to use...

10 November 2016 709 3 View

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02 March 2021 3,060 3 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 View

Can we distinguish the type of free fluid inside the pelvic and abdominal cavities whether blood or serous fluid using the ultrasound?

during ultrasound examination of acute abdomen, sometimes we see free fluid , can we know the type of fluid ,whether blood or not , using ultrasound only? if yes , what are the sonographic...

01 March 2021 7,476 4 View

Issues with Permeabilization during Flow Cytometry?

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

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Can Virtual Servers (remote dekstop manager) run heavy ANSYS Fluent CFD simulation ?

I have a virtual machine. My host Pc specs are not that good thats why i went for virtual servers. so basically my host pc has 8gb of ram with nvidia quadro k2200 4Gb DDR5 GPU ,processor intel...

01 March 2021 8,415 1 View

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

What could be some reasons why unstimulated monocytes are getting activated?

Im doing PBMC isolation -> CD14+ enrichment using magnetic beads -> stimulation setup. My negative control is just cells in cRPMI but they seem to get activated over and over again.

28 February 2021 7,883 3 View