Determining RNA quality after RNA-IP for RIP-Seq. Bioanalyzer ?

Does anyone have experience assessing RNA quality after RNA-immunoprecipitation? My downstream goal is RNA-seq (RIP-seq), and I want to check the quality of the RNA before proceeding to library...

01 February 2018 860 3 View

Tips for protein resolution: 65 vs 75 kD?

Due to limited sample, I'm trying to maximize each SDS gel/Western Blot that I run. In an ideal world, I'd like to resolve a 40 kD, 65 kD, and 75 kD protein on the same gel. I prefer to cut my...

10 November 2017 6,183 4 View

Does anyone know of a good resource for mass spec analysis workflow?

07 August 2017 6,620 9 View

My protein was detectable by Western Blot but not mass spec...ideas?

04 May 2017 1,517 5 View

Remove glycerol from antibodies?

03 April 2017 7,043 12 View

Is sonication of lysate required for crosslink-RNA-IP?

I'm doing a crosslink RNA IP. Briefly, my cells are crosslinked with formaldehyde prior to cell lysis and immunoprecipitation. After eluting my sample and reversing the crosslink, RNA is extracted...

02 March 2017 7,355 3 View

RT-qPCR: Very different amplification curves between samples. Short linear phase? Drooping plateau?

09 October 2016 8,423 7 View

Does anyone have experience with RNA-IP: enrichment over input qPCR validation?

07 August 2014 1,352 9 View

Is there any test to determine the cardiovascular risk of patients with obesity? from the perspective of obesity?

We found tests to determine cardiovascular risk taking into account lipid profile, blood pressure, age, history of diabetes, other laboratories. But when weight is involved, there is a paradox. I...

02 March 2021 6,036 1 View

No title

02 March 2021 3,060 3 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

What is the state of the drug inside solid lipid nanoparticles (SLNs)?

I am a little confuse with the mechanism of solid lipid nanoparticles (SLNs). The main point of encapsulating drugs into SLNs is because some drugs have low solubility in water. Therefore, SLNs...

01 March 2021 3,703 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

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

Interested to stain mice tissue with WGA 488 conjugate, anybody has a protocol to recommend?

Interested to stain mice brain tissue with WGA 488 conjugate, appreciate if anyone willing to offer me some guidance or suggestions! Thank you!

28 February 2021 3,951 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