EpiTect ChIP qPCR Assays

ChIP Mouse - NIH3T3

Experiment
ChIP Mouse - NIH3T3
Product
EpiTect ChIP qPCR Assays from Qiagen
Manufacturer
Qiagen

Protocol tips

Upstream tips
-Warm IP Lysis Buffer to room temperature to prevent precipitation.
-Prepare one extra plate of cells to estimate cell number.
-Thaw Protease Inhibitor Cocktail (PIC) at room temperature.
-It is recommended to take some trial to optimize sonication condition before beginning with sonication.
Protocol tips
-In general it is recommended that one million mammalian cells are required for each IP
fraction.
-Centrifuge the sample at 6000 × g for 5 min at 4 °C to eliminate bubbles.
-To reduce background, remove as much supernatant as possible in the last wash. Continue with next step OR store at 4°C overnight.

Publication protocol

Fixing and preparation for immunostaining: NIH3T3 cells were grown on rectangular patterns for 3 hr, 3 days, 6 days, and 10 days. The cells at the four-stated
time-points were fixed with 2% formaldehyde for 5 min at room temperature (RT). Following fixation, cells were quenched with 127 mM glycine for 10 min at RT. Cells were then washed with Phosphate-buffered Saline (PBS). The cells were lysed and nuclei were prepared in lysis buffer (10 mM Tris-HCl (pH 8), 10 mM NaCl, 0.2% IGEPAL CA-630 (Sigma)) with protease inhibitor cocktail (Roche) for 30 min on ice with intermittent agitation. Nuclei were washed with 1x Fast Digest (FD) buffer (Thermo Fisher Scientific). 400 μl of 1 x FDbuffer and 6 μl
of 20% SDS was added to the nuclei and incubated at 37˚C for 60 min with constant agitation.

Full paper   Login or join for free to view the full paper.

Reviews

EpiTect ChIP qPCR Assays from Qiagen has not yet been reviewed for this experiment

We'd love it if you would be the first to write a review!

Discussion

Start your discussion

Share your thoughts or question with experts in your field

Start a discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing ChIP Mouse - NIH3T3 using EpiTect ChIP qPCR Assays from Qiagen.

View full paper   Login or join for free to view the full paper.

Manufacturer protocol

Download the product protocol from Qiagen for EpiTect ChIP qPCR Assays below.

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing ChIP Mouse - NIH3T3 using EpiTect ChIP qPCR Assays from Qiagen. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.

We haven't found any additional videos for this experiment / product combination yet.

Outsource your experiment

Fill out your contact details and receive price quotes in your Inbox

  Outsource experiment
Become shareholder Discussions About us Contact Privacy Terms