Anti ddx11

Manufactured by Santa Cruz Biotechnology

Anti-DDX11 is a research antibody that specifically binds to the DDX11 protein. DDX11 is a DNA helicase that plays a role in chromosome segregation and DNA repair. This antibody can be used to detect and study the DDX11 protein in various experimental applications.

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3 protocols using anti ddx11

Antibody Staining for DNA Damage Response Proteins

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As primary antibodies, anti-DDX11 (Santa Cruz #sc-271711) (1:1,000), anti-BRCA1 (Santa Cruz #sc-6954) (1:1,000), anti-BRCA2 (ab123491) (1: 1000), anti-Chk2 (Santa Cruz #sc-17747) (1:1,000), anti-pChk2 (pT68) (cell signaling technology #2661) (1:1,000), anti-DNA PKcs (Epitomics #1579–1) (1:1,000), anti-DNA PKcs (pS2056) (Epitomics #3892–1) (1:1,000), anti–γ-H2AX (Millipore #05–636) (1:500), anti-53BP1 (Novus Biologicals #NB100-304) (1:1,000), anti-MAD2B (REV7) (BD Biosciences 612266) (1:1,000), anti-RAD51 (Santa Cruz #sc-17747) (1:50), anti-RPA2 (RPA32) (Thermo Fisher #MA1-26418), and α-tubulin (Santa Cruz #8035) (1:5,000) were used. As secondary antibodies, anti-mouse HRP-linked (1:5,000 cell signaling technology), anti-rabbit HRP-linked (1:5,000 cell signaling technology), and Alexa Fluor 488 anti-mouse (immunofluorescence 1:400) Invitrogen, Alexa Fluor Cy3-conjugated anti-rabbit (immunofluorescence 1:400) Invitrogen were used.

Jegadesan N.K, & Branzei D. (2021). DDX11 loss causes replication stress and pharmacologically exploitable DNA repair defects. Proceedings of the National Academy of Sciences of the United States of America, 118(17), e2024258118.

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DDX11 Protein Expression Analysis

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Fractionated (nucleus/cytoplasm) and not‐fractionated cell extracts were prepared from LFB cell lines using M‐PER™ Mammalian Protein Extraction Reagent (Thermo Fisher Scientific) as previously reported (Bottega et al., 2018). Primary antibodies were used as follows: anti‐DDX11 (Santa Cruz, sc‐271711, 1:500) and anti‐β‐actin (Santa Cruz, sc‐47778, 1:2000). Immuno‐reactivity was visualized using the Enhanced Chemiluminescent SuperSignal™ West Femto Maximum Sensitivity Substrate (Pierce).

Bottega R., Napolitano L.M., Carbone A., Cappelli E., Corsolini F., Onesti S., Savoia A., Gasparini P, & Faletra F. (2019). Two further patients with Warsaw breakage syndrome. Is a mild phenotype possible?. Molecular Genetics & Genomic Medicine, 7(5), e639.

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Protein Isolation and Fractionation from LCLs

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Protein whole and fractionated cell extracts were prepared from patients' lymphoblastoid cell lines (LCLs) as previously described (Bottega et al., 2018) . Briefly, M-PER™ Mammalian Protein Extraction Reagent and NE-PER™ Nuclear and Cytoplasmic Extraction Reagents (Thermo Fisher Scientific), respectively, were used. Primary antibodies used were anti-DDX11 (sc-271711; Santa Cruz), anti-Sp1 (sc-17824; Santa Cruz), and anti-Hsp90 (sc-13119; Santa Cruz,).
Immunoreactivity was visualized using the Enhanced Chemiluminescent SuperSignal™ West Femto Maximum Sensitivity Substrate (Pierce). The protein amount in each subcellular fraction was calculated by densitometric analysis and normalized with respect to the total amount of DDX11.

Bottega R., Ravera S., Napolitano L.M.R., Chiappetta V., Zini N., Crescenzi B., Arniani S., Faleschini M., Cortone G., Faletra F., Medagli B., Sirchia F., Moretti M., de Lange J., Cappelli E., Mecucci C., Onesti S., Pisani F.M, & Savoia A. (2021). Genomic integrity and mitochondrial metabolism defects in Warsaw syndrome cells: a comparison with Fanconi anemia. Journal of cellular physiology, 236(8).

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