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The RPA32 is a laboratory equipment designed for the rapid amplification of nucleic acids. It utilizes the Reverse Transcription Polymerase Chain Reaction (RT-PCR) method to quickly and accurately detect and quantify target genetic sequences.

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Lab products found in correlation

Proliferating cell nuclear antigen (pcna), by Santa Cruz Biotechnology (2 mentions) Rpa32, by Fortis Life Sciences (1 mentions) Topbp1, by Santa Cruz Biotechnology (1 mentions) γh2ax, by Cell Signaling Technology (1 mentions) Xrcc4, by Thermo Fisher Scientific (1 mentions) Flag tag (flag), by Merck Group (1 mentions) Gapdh, by Merck Group (1 mentions) γh2ax, by Merck Group (1 mentions) β actin, by Fortis Life Sciences (1 mentions) Rad51, by Santa Cruz Biotechnology (1 mentions) Prpas33, by Merck Group (1 mentions) Gh2ax, by Merck Group (1 mentions) Axiocam 506 color camera, by Zeiss (1 mentions)

4 protocols using rpa32

1

Immunoblotting Analysis of DNA Damage Response

Cited 2 times
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Immunoblotting analysis was performed as described previously (27–29 (link),60 (link),64 (link)). Primary antibodies were purchased from respective vendors: APE1 (Santa Cruz Biotechnology Cat#sc-17774), ATR (Santa Cruz Biotechnology Cat#515173), ATM (GeneTex Cat#GTX70103), ATM phosphorylation at Ser1981 (Abcam Cat#ab81292), ATR phosphorylation at Thr1989 (Cell Signaling Technology Cat#D5K8W), ATRIP (Santa Cruz Biotechnology Cat#365383), Chk1 (Santa Cruz Biotechnology Cat#sc-8408), Chk1 phosphorylation at Ser345 (Cell Signaling Technology Cat#133D3), Chk1 phosphorylation at Ser317 (Cell Signaling Technology Cat#D12H3), Chk2 (Santa Cruz Biotechnology Cat#sc-9064), Chk2 phosphorylation at Thr68 (Santa Cruz Biotechnology Cat#sc-16297), eGFP (Life Technologies Corporation Cat#TA150041), ETAA1 (Abcam Cat#ab122245), H2AX (Cell Signaling Technology Cat#2D17A3), H2AX phosphorylation at Ser139 (Cell Signaling Technology Cat#2577s), NPM1 (Santa Cruz Biotechnology Cat#sc-47725), p53 phosphorylation at Ser15 (Cell Signaling Technology Cat#9284), p53 (Santa Cruz Biotechnology Cat#sc-126), PCNA (Santa Cruz Biotechnology Cat#sc-56), RPA32 (Thermo Fisher Scientific Cat#MA1-26418), RPA32 phosphorylation at Ser33 (Bethyl Laboratories Cat#A300-246A), TopBP1 (Santa Cruz Biotechnology Cat#271043), Tubulin (Santa Cruz Biotechnology Cat#sc-8035), YFP (BioVision Cat#3991–100).
Li J., Zhao H., McMahon A, & Yan S. (2022). APE1 assembles biomolecular condensates to promote the ATR–Chk1 DNA damage response in nucleolus. Nucleic Acids Research, 50(18), 10503-10525.
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2

DNA Damage Response Protein Profiling

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Antibodies used were: RAD51AP1 (Proteintech), XRCC4 (Thermo Fisher Scientific), RAD51 (Santa Cruz), γH2AX (Millipore), γH2AX (Cell Signaling), RPA32 (Thermo Fisher Scientific), PCNA (Santa Cruz), Flag (Sigma), S9.6 (Millipore), mouse or rabbit control IgG (Diagenode), β-actin (Bethyl), GAPDH (Millipore).
Ouyang J., Yadav T., Zhang J.M., Yang H., Rheinbay E., Guo H., Haber D.A., Lan L, & Zou L. (2021). RNA transcripts stimulate homologous recombination by forming DR-loops. Nature, 594(7862), 283-288.
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3

Immunoblotting Analysis of Cell Lysates

Cited 2 times
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Immunoblotting analysis of cell lysates or Xenopus egg extracts was carried out similarly as we described previously (Willis et al., 2013 (link); Wallace et al., 2017 (link); Lin et al., 2018 (link), 2020 (link)). Primary antibodies against Chk1 (Santa Cruz Biotechnology Cat#sc-8408), Chk1 phosphorylation Ser345 (Cell Signaling Technology Cat#133D3), RPA32 (Thermo Fisher Scientific Cat#MA1-26418), RPA32 phosphorylation Ser33 (Bethyl Laboratories Cat#A300-246A), and Tubulin (Santa Cruz Biotechnology Cat#sc-8035) were purchased from various vendors. Anti-human APE2 antibodies were prepared as described previously (Tsuchimoto et al., 2001 (link)).
Hossain M.A., Lin Y., Driscoll G., Li J., McMahon A., Matos J., Zhao H., Tsuchimoto D., Nakabeppu Y., Zhao J, & Yan S. (2021). APE2 Is a General Regulator of the ATR-Chk1 DNA Damage Response Pathway to Maintain Genome Integrity in Pancreatic Cancer Cells. Frontiers in Cell and Developmental Biology, 9, 738502.
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4

Immunofluorescence Staining of DNA Repair Proteins

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Cells on coverslips were washed with PBS and fixed in 4% PFA/2% sucrose solution for 15 min. The coverslips were washed again with PBS and then Triton extracted (0.5% Triton X-100 in PBS) for 4 min. Cells were incubated with their respective antibodies for 30 min at 37°C followed by incubation with secondary antibodies (FITC or Rhodamine) for 30 min at 37°C. Primary antibodies used in IF studies were RPA34 (Cal Biochem; NA18; 1:100), 53BP1 (Bethyl; A300-272A; 1:2,000), g-H2AX (Millipore; 05–636; 1:5,000), RPA32 (Thermo Fisher Scientific; PA5-22256; 1:400), pRPAS33 (Sigma; PLA0210-100 μl; 1:1,500), and BRCA1 (Upstate; 07–434; 1:400). Coverslips were mounted using mounting medium (DAPI). Images were acquired with an Axio Imager.M2 (Carl Zeiss) equipped with an Axiocam 506 color camera, controlled by Zen software.
Duan H., Mansour S., Reed R., Gillis M.K., Parent B., Liu B., Sztupinszki Z., Birkbak N., Szallasi Z., Elia A.E., Garber J.E, & Pathania S. (2020). E3 ligase RFWD3 is a novel modulator of stalled fork stability in BRCA2-deficient cells. The Journal of Cell Biology, 219(6), e201908192.
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