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Cdc45

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CDC45 is a protein involved in the initiation and regulation of DNA replication. It is a critical component of the pre-replication complex and plays a key role in the unwinding of DNA during the replication process.

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Lab products found in correlation

Proliferating cell nuclear antigen (pcna), by Santa Cruz Biotechnology (2 mentions) Flag tag (flag), by Merck Group (2 mentions) Actinin, by Santa Cruz Biotechnology (1 mentions) α tubulin, by Merck Group (1 mentions) Nbp1 89091, by Novus Biologicals (1 mentions) Anti rabbit hrp 32460, by Thermo Fisher Scientific (1 mentions) Anti goat hrp, by Thermo Fisher Scientific (1 mentions) Ha tag, by Cell Signaling Technology (1 mentions) Anti mouse hrp, by Thermo Fisher Scientific (1 mentions) Claspin, by Fortis Life Sciences (1 mentions) Chk1 ps317, by Cell Signaling Technology (1 mentions) Pold3, by Fortis Life Sciences (1 mentions) P chk1, by Cell Signaling Technology (1 mentions) Gapdh, by Santa Cruz Biotechnology (1 mentions) P histone h2a x s139, by Cell Signaling Technology (1 mentions) Cyclin d1, by Cell Signaling Technology (1 mentions) Geminin, by Abcam (1 mentions) P rb ser807 811, by Cell Signaling Technology (1 mentions) Fl 393, by Santa Cruz Biotechnology (1 mentions) H3k9 3me, by Merck Group (1 mentions)

4 protocols using cdc45

1

Immunoblotting of Replication Proteins

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Primary: MCM3 (sc-390480; Santa Cruz), MCM4 (A300-125A; Bethyl Laboratories), MCM6 (611622; BD Biosciences), MCM7 (sc-22782; Santa Cruz), CDC45 (sc-55569; Santa Cruz), Actinin (sc-17829; Santa Cruz), OBFC1 (STN1) (ab119263; Abcam), TEN1 (Kasbek et al, 2013 (link)), WHDH1 (AND-1) (NBP1-89091; Novus), PolA1 (A302-850A; Bethyl Laboratories), α-Tubulin (T9026; Sigma-Aldrich), H3 (9715; Cell Signaling), HA-tag ([anti-mouse: 2367; Cell Signaling] [anti-rabbit: 3724; Cell Signaling]), Flag-tag ([anti-mouse: F1804; Sigma-Aldrich] [anti-rabbit: PA1984B; Thermo Fisher Scientific]), and Myc-tag ([anti-mouse: 05-724; EMD Millipore] [anti-rabbit: ab1906; Abcam]). Secondary: Thermo Fisher Scientific: anti-rabbit-HRP (32460), anti-mouse-HRP (32430), and anti-goat-HRP (31402); molecular probes: goat–anti-mouse Alexa Fluor 647 (A21235), goat–anti-rabbit Alexa Fluor 647 (A21244), goat–anti-rabbit Alexa Fluor 594 (A11037), goat–anti-mouse Alexa Fluor 594 (A11032), goat–anti-mouse Alexa Fluor 488 (A11029), and goat–anti-rabbit Alexa Fluor 488 (A11034).
Wang Y., Brady K.S., Caiello B.P., Ackerson S.M, & Stewart J.A. (2019). Human CST suppresses origin licensing and promotes AND-1/Ctf4 chromatin association. Life Science Alliance, 2(2), e201800270.
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2

Replication Stress Response Profiling

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The following compounds were used: hydroxyurea (HU, 3mM), ATR inhibitor VE821 (ATRi, 3μM), aphidicolin (2.95μM), CDC7 inhibitor (CDC7i, 20μM) and roscovitine (20μM). HU and aphidicolin were used at 5mM and 14.75μM respectively for high dose replication stress. Antibodies used were CHK1 total (Santa Cruz), CHK1 pS317 (Cell Signaling), MCM2 (BD Biosciences), POLD3 (Bethyl), RPA2 (Abcam), PCNA (Santa Cruz), CDC45 (Santa Cruz), CLASPIN (Bethyl) and ORC2 (BD Biosciences).
Dungrawala H., Rose K.L., Bhat K.P., Mohni K.N., Glick G.G., Couch F.B, & Cortez D. (2015). The replication checkpoint prevents two types of fork collapse without regulating replisome stability. Molecular cell, 59(6), 998-1010.
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3

Replication Protein Abundance Assay

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POLE1 (Santa Cruz, #sc-390785, 1:500), osTIR1 (MBL International, #PD048, 1:1000), GAPDH (Santa Cruz, #sc-47724, 1:1000), pCHK1 (Cell Signaling, #2360S, 1:1000), Chk1 (Cell Signaling, #2348S, 1:1000), MCM4 (Cell Signaling, #3228S, 1:300), CDC45 (Santa Cruz, #sc-55569, 1:500), SLD5 (Santa Cruz, #sc‐398784, 1:300), H3 (Santa Cruz, #sc-517576, 1:1000), MCM7 (Santa Cruz, #sc-9966, 1:1000), POLE2 (Santa Cruz, #sc-398582, 1:500), PCNA (Santa Cruz, #sc-56, 1:1000), FLAG (Sigma, F3165-1MG, 1:3000), myc (Cell Signaling, #2276, 1:1000).
Vipat S., Gupta D., Jonchhe S., Anderspuk H., Rothenberg E, & Moiseeva T.N. (2022). The non-catalytic role of DNA polymerase epsilon in replication initiation in human cells. Nature Communications, 13, 7099.
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4

Immunofluorescence Analysis of DNA Damage

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The primary antibodies used were: for total p53, p53 DO-1, Santa Cruz sc-126 (Dallas, TX, USA) and FL-393 Santa Cruz; p53 (S15 phosphorylation, Cell Signaling 9286S, Boston, MA, USA); p-Histone H2A.X (S139) (Cell Signaling 2577S); RPA32 (4E4 Cell Signaling 2208S); BrdU (BD Pharmingen 555627, BD Biosciences, San Jose, CA, USA); cleaved Caspase 3 (Asp 173, Cell Signaling 9661S); Cyclin E (Abcam ab3927, Cambridge, UK); PCNA (Abcam ab18197); MCM4 (Abcam ab4459); MCM5 (Abcam ab17967); Orc1 (Abcam ab 85830); Cdt1 (Abcam ab70829); Geminin (Abcam ab12147); β-Actin (Sigma A1978); pRb Ser 807/811 (Cell Signaling 9308S); p21 (2946 Cell Signaling); Cyclin D1 (2926 Cell Signaling); cdc45 (Santa Cruz sc-20685); Chk1 (Santa Cruz sc-8408); Chk1 pS345 (Cell signalling 2341) H3K9-3Me (Millipore 17–625, Billerica, MA, USA).
Maya-Mendoza A., Merchut-Maya J.M., Bartkova J., Bartek J., Streuli C.H, & Jackson D.A. (2014). Immortalised breast epithelia survive prolonged DNA replication stress and return to cycle from a senescent-like state. Cell Death & Disease, 5(7), e1351-.
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