Samantha Jeschonek

9 Questions 54 Answers 0 Followers

Questions related from Samantha Jeschonek

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...

02 February 2018 940 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...

11 November 2017 6,259 4 View

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

Hello all -- Does anyone have a good paper/book/website that goes through the analysis of (label-free) mass spec data starting from .raw files? I'm specifically looking for resources that use...

08 August 2017 6,720 9 View

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

We did an IP experiment and sent the results for mass spec. I confirmed by western blot (on the same sample) that some positive controls were present -- including the antigen. The mass spec...

05 May 2017 1,592 5 View

Does anyone have a protocol for coating microscope slides with lipids?

Hi all -- We are hoping to coat microscope slides in the lipid DOPC. Does anyone have protocol for this?

05 May 2017 3,864 1 View

Remove glycerol from antibodies?

Can anyone recommend a method or product to remove glycerol from small volumes of affinity purified antibody? We are using Dynabead M270 Epoxy beads to couple antibodies to beads. This kit...

04 April 2017 7,130 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...

03 March 2017 7,433 3 View

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

I'm having some trouble with my RT-qPCR. Some of my amplification curves (delta RN vs cycle #) look as expected, but others have a distorted curve. High starting background that drops, followed...

10 October 2016 8,523 7 View

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

Hello - I have a question regarding the best way to setup a (two step) RT-qPCR experiment. I have performed an RNA-IP and using qPCR want to determine if a specific RNA is enriched over (1) input...

08 August 2014 1,420 9 View