Human Perforin Antibody Summary
Accession # P14222
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Detection of Human Perforin by Western Blot. Western blot shows lysates of human NK cells and HDLM‑2 human Hodgkin's lymphoma cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human Perforin Monoclonal Antibody (Catalog # MAB8011) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Perforin at approximately 70 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Perforin in Human Spleen. Perforin was detected in immersion fixed paraffin-embedded sections of human spleen using Mouse Anti-Human Perforin Monoclonal Antibody (Catalog # MAB8011) at 5 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 Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell surfaces and cytoplasm in splenocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Perforin is a pore forming, cytolytic protein that is a component of cytolytic T-cell and NK cell granules. Perforin activates clathrin- and dynamin-dependent endocytosis, which removes perforin and granzymes from the plasm membrane to early endosomes, preserving outer membrane integrity. Perforin facilitates granzyme uptake and avoids the proinflammatory necrotic death of a membrane-damaged cell. Defects in the Perforin gene results in Familial hemophagocytic lymphohistiocytosis (FHL). Perforin-based effector systems are involved not only in the lysis of abnormal cells but also in the down-regulation of cellular immune activation.
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