LaCava Research Wiki

First Jess experiment was to test anti-rrp6 Ab. Basically, I tried to confirm my western blots with Jess. https://macromolecule-child.rockefeller.edu/#Oct%202020%20HEK%20293%20RRP6-3xFlag%20Flp-In%20TREX%20cell%20line%20DOX%20test%20induction

There lines without lysate to check Ab off-targets, lines without 1st Ab, lines with different concentrations of 1st Ab - 1:10, 1:50 and 1:250. For next experiment I decided to use 1:250 of anti-rrp6 Ab.

Normally we use anti-rrp6 Ab in 1:1000 concentration, so using 4x (1:250), 20x (1:50) and 100x (1:10) was recommended by Jess company to test Ab saturation.

As a secondary Ab I used anti-rabbit secondary provided by company

As a protein sample (lysate) here I used HEK 293 parental cell line. So it should show only one band of rrp6 protein ~ 100 kDa.

I also tested different concentrations of lysate - 0.2 , 0.4 and 1 ug/ul.

For next experiment I used sample concentration 0.4 ug/ul

separation time 25 min (normal settings)

report:

http://ruselfhost.rockefeller.edu/WWW/sites/macromolecule-child.rockefeller.edu/tiddlywiki/scientific/Nastya_data/Jess/anti_rrp6_ab_test/Nastya_Jess_anti_rrp6_antibody_test%20%202020-10-14_12-09-18_Jess.cbz

^This is .cbz fIle with all info. Need to be opened in Compass for SW software

PDF report: