Rabbit anti gfp

Primary antibodies used were rabbit anti‐p65 from Santa Cruz (clone C‐20, ref. sc‐372) at 1/250, mouse anti‐myc9E10 (developed by Bishop, J.M. was obtained from the Developmental Studies Hybridoma Bank, created by the NICHD of the NIH and maintained at the University of Iowa), mouse anti‐HA (Eurogentec, clone 16B12, ref. MMS‐101R), rabbit anti‐HA (Sigma, ref. H6908), rabbit anti‐GFP (Amsbio, ref. TP401), rabbit anti‐UBAP1 (Proteintech, ref. 12385‐1‐AP), mouse anti‐His (Sigma, clone HIS‐1, ref. H1029), mouse anti‐FLAG (Sigma, clone M2, ref. F1804) all at 1/1000 and mouse anti‐TNFR1 (Santa Cruz, clone H‐5, ref. sc‐8436) and rabbit anti‐TSG101 (Atlas Antibodies, ref. HPA006161) both at 1/200. Rabbit polyclonal anti‐PumA serum was obtained by repeated immunization of rabbits with purified PumA (Eurogentec) and was used at 1/1,000 for Western blot and for immunofluorescence microscopy. Purified BtpA was used to obtain chicken anti‐BtpA (Eurogentec). Anti‐EF‐Tu antibody (kind gift from R. Voulhoux) was used at 1/10,000.
Secondary antibodies used were anti‐rabbit, mouse, chicken or rat conjugated with Alexas‐488, ‐555 or ‐647 all from Jackson ImmunoResearch. When necessary, phalloidin‐568 (1/1,000) was used to label the actin cytoskeleton and DAPI nuclear dye (1/1,000) for the host cell nucleus. For Western blots, anti‐mouse or rabbit‐HRP antibodies were used at 1/5,000.

For western blot analysis, cells were fixed with 20% trichloroacetic acid (TCA) and disrupted by bead beating. Lysate and precipitate/debris were mixed with 600 µL 10% TCA and pelleted. The pellet was resuspended in 1× Laemmli buffer with 5% β-mercaptoethanol and 160 mM Tris-HCl (pH 7.4), boiled for 10 min and sonicated briefly. The extract was directly subjected to sodium dodecyl sulfate (SDS) gel analysis without clearance. The following antibodies were used: mouse monoclonal anti-Rad53 (clone EL7, in house^{58 (link)}, 1:5), mouse monoclonal anti c-MYC (clone 9E10, Santa Cruz Biotechnology Cat# sc-40, RRID:AB_627268, 1:2000), rabbit anti-histone H3 (EpiCypher, Cat# 13-0001, 1:5000), rabbit anti-histone H4 (Abcam Cat# 7311, 1:4000), rabbit anti-histone H3 (acetyl K27) (Abcam, Cat# ab4729, 1:2000), mouse monoclonal anti-acetyl-lysine (T52, in-house^{59 (link)}, 1:10), mouse monoclonal anti-Pgk1 (novex, Cat# 459250, 1:10,000), mouse monoclonal anti-Porin (abcam, Cat# 110326, 1:1000), rabbit anti-GFP (Amsbio, Cat# TP401, 1:5000), goat anti-mouse IgG (H + L)-horseradish peroxidase (HRP) Conjugate (Bio-Rad, Cat# 1706516, 1:20,000), goat anti-rabbit IgG (H + L)-HRP Conjugate (Bio-Rad, Cat# 1706515, 1:20,000). Detection was performed by electrochemiluminescence (GE Healthcare). Uncropped western blots are shown in Supplementary Figs. 10 and 11.