Pold3

Manufactured by Fortis Life Sciences

POLD3 is a laboratory equipment product that functions as a DNA polymerase subunit. It is used in various molecular biology applications involving DNA synthesis and amplification.

Automatically generated - may contain errors

Lab products found in correlation

Rnf168, by Merck Group (1 mentions) Phospho chk1, by Cell Signaling Technology (1 mentions) Ne per nuclear and cytoplasmic extraction kit, by Thermo Fisher Scientific (1 mentions) Flag tag (flag), by Cell Signaling Technology (1 mentions) Palb2, by Fortis Life Sciences (1 mentions) Brca2, by Fortis Life Sciences (1 mentions) Rad18, by Fortis Life Sciences (1 mentions) Rad51, by Santa Cruz Biotechnology (1 mentions) Proliferating cell nuclear antigen (pcna), by Santa Cruz Biotechnology (1 mentions) Claspin, by Fortis Life Sciences (1 mentions) Cdc45, by Santa Cruz Biotechnology (1 mentions) Chk1 ps317, by Cell Signaling Technology (1 mentions)

2 protocols using pold3

Western Blot Analysis of DNA Damage Response Proteins

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Nuclear extracts were obtained using the NE-PER Nuclear and Cytoplasmic Extraction Kit (Thermo Scientific) and whole cell extracts were generated using RIPA buffer with protease and phosphatase inhibitors added. Proteins were separated by sodium dodecyl sulphate (SDS)-polyacrylamide gelelectrophoresis and transferred to a polyvinylidene fluoride (PVDF) membrane. Membranes were blocked with 5% nonfat milk in phosphate-buffered saline tween 20 (PBST) at room temperature for 1 h. Primary antibodies were incubated overnight at 4° and horseradish peroxidase (HRP)-conjugated secondary antibodies were incubated for 1 h at room temperature. The following primary antibodies were used: BRCA1 (EMD Millipore, catalog# OP92), RNF168 (EMD Millipore, #06-1130-I), Tubulin (Cell Signaling, catalog# 2148), GFP (Santa Cruz Biotechnology, catalog# sc-9996), RFP (ChromoTek, catalog# 6g6-20), FLAG (Cell Signaling, catalog# 14793 and Sigma Aldrich, catalog# F1804), phospho-Chk1 (Cell Signaling, catalog# 2344), Chk1 (Cell Signaling, catalog# 2360), POLD3 (Bethyl Laboratories, A301-244A), PALB2 (Bethyl Laboratories, catalog# A301-246A), BRCA2 (Bethyl Laboratories, catalog# A303-435A), RAD51 (Santa Cruz Biotechnology, catalog# sc-8349), RAD18 (Bethyl Laboratories, catalog# A301-340A), 53BP1 (Cell Signaling, catalog# 4908).

Krais J.J, & Johnson N. (2020). Ectopic RNF168 expression promotes break-induced replication-like DNA synthesis at stalled replication forks. Nucleic Acids Research, 48(8), 4298-4308.

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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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