PARP1 IHC Staining Protocol

PARP1 IHC staining was carried out using a primary polyclonal rabbit anti-PARP1 antibody (sc-7150, 0.4 μg/mL, Santa Cruz Biotechnology). At MSK, the staining was carried out using the automated Discovery XT processor (Ventana Medical Systems) at the Molecular Cytology Core Facility as described previously^{26 (link)}. At MSMC, the same anti-PARP1 antibody was used (0.4 μg/mL, 1 h) in combination with the DAKO Real Envision kit (K5007, DAKO). Since the PARP1 antibody was discontinued by Santa Cruz, we tested and validated PA5–16452 (Invitrogen) as potential alternative.

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Kossatz S., Pirovano G., De Souza França P.D., Strome A.L., Sunny S.P., Zanoni D.K., Mauguen A., Carney B., Brand C., Shah V., Ramanajinappa R.D., Hedne N., Birur P., Sihag S., Ghossein R.A., Gönen M., Strome M., Suresh A., Molena D., Ganly I., Kuriakose M.A., Patel S.G, & Reiner T. (2020). Validation of the use of a fluorescent PARP1 inhibitor for the detection of oral, oropharyngeal and oesophageal epithelial cancers. Nature biomedical engineering, 4(3), 272-285.

Publication 2020

sc-7150 Discovery XT processor DAKO Real Envision kit PA5–16452

Anti antibody Antibody Cytology Parp1 Rabbit

Corresponding Organization : Cornell University

Other organizations : Memorial Sloan Kettering Cancer Center, Narayana Health, Mazumdar Shaw Medical Foundation, Anesthesiology and Surgical Oncology Research Group

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Variable analysis

independent variables

Primary antibody used (anti-PARP1 antibody from Santa Cruz Biotechnology vs. Invitrogen)
Staining protocol (automated Discovery XT processor vs. DAKO Real Envision kit)

dependent variables

PARP1 IHC staining

control variables

Antibody concentration (0.4 μg/mL)
Antibody incubation time (1 hour)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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