α-PCNA (P8825) and α-HIS (H1029) were purchased from Sigma; α-Cul2 (EPR3104) was purchased from Abcam; α-p97 (65278) was purchased from Progen Biotechnik, anti-P-Chk1 (S345) from Cell Signaling, γ-H2AX (4418-APC-020, Trevigen).
Affinity purified α-Cdc45, α-Psf2 (40 (link)), α-Mcm3 (41 (link)), α-LRR1 (S962D) and α-Cul2 (SA206) (4 (link)), and α-Mcm7 (6 ) were previously described. Xenopus Ufd1 antibody was a kind gift from Prof Stemmann’s lab (13 (link)).
Xenopus full-length p97, Ubxn7, Faf1, and GINS proteins were purified as described previously (6 , 13 (link)) and antibodies raised against such prepared antigens in sheep (p97, GINS, Ubxn7) or rabbit (Faf1). The resulting antibody sera were purified in-house against the purified antigen. The specificity of each new antibody is presented inFig S7 .
Affinity purified α-Cdc45, α-Psf2 (40 (link)), α-Mcm3 (41 (link)), α-LRR1 (S962D) and α-Cul2 (SA206) (4 (link)), and α-Mcm7 (6 ) were previously described. Xenopus Ufd1 antibody was a kind gift from Prof Stemmann’s lab (13 (link)).
Xenopus full-length p97, Ubxn7, Faf1, and GINS proteins were purified as described previously (6 , 13 (link)) and antibodies raised against such prepared antigens in sheep (p97, GINS, Ubxn7) or rabbit (Faf1). The resulting antibody sera were purified in-house against the purified antigen. The specificity of each new antibody is presented in
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