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Mcm7 sc 9966

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MCM7 (sc-9966) is a primary antibody that recognizes the MCM7 protein. MCM7 is a component of the MCM complex, which is involved in the initiation and regulation of eukaryotic genome replication.

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Lab products found in correlation

Mcm3 a300 192a, by Fortis Life Sciences (2 mentions) Anti pcna sc 56, by Santa Cruz Biotechnology (2 mentions) Anti flag antibody f1804 200ug, by Merck Group (2 mentions) Set8 14063 1 ap, by Proteintech (2 mentions) Anti ddb2 pa5 79143 and pa5 63568, by Thermo Fisher Scientific (2 mentions) Anti histone h3 antibody ab1791, by Abcam (2 mentions) Phospho chk1 ser345, by Cell Signaling Technology (1 mentions) Laminb1 ab16048, by Abcam (1 mentions) Pd166285, by Merck Group (1 mentions) Nms 873, by Merck Group (1 mentions) Confocal microscope, by Leica (1 mentions) Rpa32 pt21, by Abcam (1 mentions) Rb sc 102, by Santa Cruz Biotechnology (1 mentions)

Spelling variants of this product

Sc-9966 (2 citations)

5 protocols using mcm7 sc 9966

1

Antibody Characterization for Cell Cycle

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Anti-CDT1 (A300-786A), CDT2 (A300-947A), MCM2 (A300-191A) and MCM3 (A300-192A) antibodies were purchased from Bethyl Laboratories Inc. Anti-Histone H3 antibody (ab1791) was from Abcam, and anti-Flag antibody (F1804-200UG) was from Sigma-aldrich. Anti-GAPDH (60004-1-Ig), HA (66006-2-Ig), p21 (10355-1-AP) and SET8 (14063-1-AP) antibodies were purchased from Proteintech. Anti-PCNA (sc-56) and MCM7 (sc-9966) antibodies were purchased from Santa Cruz Biotechnologies. Anti-DDB2 (PA5-79143 and PA5-63568) antibodies were from Invitrogen. Homemade polyclonal antibodies of CDT2 and CUL 1 were gifts from Dr. Hui Zhang (Department of Chemistry and Biochemistry, University of Nevada, Las Vegas, Nevada 89154, USA).
Wu X., Yu M., Zhang Z., Leng F., Ma Y., Xie N, & Lu F. (2021). DDB2 regulates DNA replication through PCNA-independent degradation of CDT2. Cell & Bioscience, 11, 34.
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2

Cell Cycle Regulation Antibodies and Inhibitors

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Antibodies to Psf2, XCAP-E, claspin, and Cut5/TopBP1 were prepared as per directions (23 (link), 52 (link), 53 (link)). Antisera to Cdc45 and Polε (p60) were provided by H. Takisawa and Y. Kubota (Osaka University). Antiserum to Polδ (p66) was provided by S. Waga (Japan Women’s University). Antibodies to APC3 and Cyclin B2 were provided by S. Mochida (Kumamoto University). The following antibodies were obtained from the indicated companies: MCM7 (sc-9966, Santa Cruz), MCM4 (ab4459, Abcam), lamin B1 (ab16048, Abcam), β-actin (ab8224, Abcam), Phospho-Chk1 (Ser345) (2341, Cell Signaling), Phospho-CDK1 (Thr14, Tyr15) (44-686G, Thermo Fisher). Recombinant His-p27 and His-geminin were prepared as described (50 (link)) and used at 100 μg/ml and 50 μg/ml to inhibit CDK activities and replication licensing, respectively. NMS-873 (Sigma-Aldrich) and PD166285 (Sigma-Aldrich) were used at 100 μM and 10 μM to inhibit p97/VCP and Wee1/Myt1 kinase activities, respectively.
Hashimoto Y, & Tanaka H. (2020). Ongoing replication forks delay the nuclear envelope breakdown upon mitotic entry. The Journal of Biological Chemistry, 296, 100033.
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3

Immunofluorescence Staining of DNA Damage Markers

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Cells were seeded on glass coverslips and treated with drug (AZD7762, CHIR-124, MK1775) for 48 hours. Cells were washed in PBS and fixed in 4% paraformaldehyde. Then they were permeabilized in 0.5% Triton X-100. For PCNA staining, cells were pre-extracted with CSK buffer (10mM HEPES, 300mM Sucrose, 100mM NaCl, and 3mM MgCl2) and extraction buffer (50mM NaF, 0.1mM NaOrthovanadate, 1mM PMSF, 0.5% Triton X-100, protease inhibitor). After permeabilization, cells were blocked in IF buffer (5% BSA, 0.4% NP40 in PBS) and incubated with primary antibody diluted in IF buffer for one hour in 37°C. Primary antibodies used in this study were: yH2AX (phospho-ser139) from Cell Signaling, MCM7 (sc-9966) from Santa Cruz, Anti-phospho-Histone H3 (ser10) (06-570) from Millipore, and RPA32 (pT21) from Abcam. Coverslips were washed in PBS and secondary antibody diluted in IF buffer was applied for one hour in 37°C. Coverslips were washed again after secondary and mounted on slides. Images were taken with either Leica Confocal Microscope at 63× magnification or fluorescence microscope at 40×. Quantification was performed using ImageJ.
Chung S., Vail P., Witkiewicz A.K, & Knudsen E.S. (2018). Coordinately targeting cell cycle checkpoint functions in integrated models of pancreatic cancer. Clinical cancer research : an official journal of the American Association for Cancer Research, 25(7), 2290-2304.
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4

Immunohistochemical Analysis of Lung Cancer

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This work was approved by the Ethics Review Committee of the First Affiliated Hospital of Xi’an Jiaotong University. A total of 147 lung cancer specimens and adjacent normal tissues were obtained from the First Affiliated Hospital of Xi’an Jiaotong University. After antigen retrieval in sodium citrate buffer (PH = 9.0) in a microwave oven, the paraffin‐embedded tissues were stained with Spt16 (8D2, Biolegend), MCM7 (sc‐9966, Santa Cruz), and Rb (sc‐102, Santa Cruz) antibodies following the manufacturer’s protocol for the LSAB detection kit (ZSGB‐BIO). Each section was scored by the staining intensity (0, none; 1, weak; 2, moderate; and 3, strong) and the positive cell rate (0, no positive cells; 1, 10%–25%; 2, 25%–50%; 3, 50%–75%; 4, >75%). The total score is the product of the positive cell rate by staining intensity.
Yang L., Wang X., Jiao X., Tian B., Zhang M., Zhou C., Wang R., Chen H., Wang B., Li J., Liu J., Zhang G, & Liu P. (2020). Suppressor of Ty 16 promotes lung cancer malignancy and is negatively regulated by miR‐1227‐5p. Cancer Science, 111(11), 4075-4087.
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5

Antibody Characterization for Cell Cycle

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Anti-CDT1 (A300-786A), CDT2 (A300-947A), MCM2 (A300-191A) and MCM3 (A300-192A) antibodies were purchased from Bethyl Laboratories Inc. Anti-Histone H3 antibody (ab1791) was from Abcam, and anti-Flag antibody (F1804-200UG) was from Sigma-aldrich. Anti-GAPDH (60004-1-Ig), HA (66006-2-Ig), p21 (10355-1-AP) and SET8 (14063-1-AP) antibodies were purchased from Proteintech. Anti-PCNA (sc-56) and MCM7 (sc-9966) antibodies were purchased from Santa Cruz Biotechnologies. Anti-DDB2 (PA5-79143 and PA5-63568) antibodies were from Invitrogen. Homemade polyclonal antibodies of CDT2 and CUL 1 were gifts from Dr. Hui Zhang (Department of Chemistry and Biochemistry, University of Nevada, Las Vegas, Nevada 89154, USA).
Wu X., Yu M., Zhang Z., Leng F., Ma Y., Xie N., & Lu F. (2020). DDB2 Regulates DNA Replication through PCNA-Independent Degradation of CDT2.
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