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Detection of PAR in Jurkat cells by Flow Cytometry.

Jurkat human acute T cell leukemia cell line treated with 1 mM H₂O₂ for 5 minutes (filled histogram) or resting (open histogram) were stained with Mouse Anti-PAR Monoclonal Antibody (Catalog # 4335-MC-100) followed by Goat anti-Mouse IgG PE-conjugated Secondary Antibody (F0102B). To facilitate intracellular staining, cells were fixed and permeabilized using FlowX FoxP3/Transcription Factor Fixation & Perm Buffer Kit (FC012). Staining was performed using our Staining Intracellular Molecules protocol.

Detection of PAR/pADPr in Human Kidney.

PAR/pADPr was detected in immersion fixed paraffin-embedded sections of human kidney using Mouse Anti-PAR/pADPr Monoclonal Antibody (Catalog # 4335-MC-100) at 1.7 µg/ml for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to the nucleus. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of PAR/pADPr in 786-O Human Cell Line.

PAR/pADPr was detected in immersion fixed 786-O human renal cell adenocarcinoma cell line untreated (negative) or treated (positive) with 2mM hydrogen peroxide for 5 minutes using Mouse Anti-PAR/pADPr Monoclonal Antibody (Catalog # 4335-MC-100) at 3 µg/ml for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to the nucleus upon stimulation with hydrogen peroxide. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.