Perforin Monoclonal Antibody (eBioOMAK-D), eBioscience™, Invitrogen™

Description

• Description: The eBioOMAK-D antibody reacts with mouse perforin (pore-forming protein, pfp, Prf)
• Perforin is one of the cytolytic mediators present in the cytoplasmic granules of cytotoxic T lymphocytes (CTL) and natural killer cells (NK)
• Perforin is involved in the killing function by CTLs and NKs and has an important role in the immune response against tumors and virus infections
• By immunobloting, eBioOMAK-D recognizes a ∽70kDa band in lysates of CTLL-2 mouse cytotoxic cell line and in lysates of IL-2 stimulated but not unstimulated mouse splenocytes
• By multi-color intracellular flow cytometric analysis, eBioOMAK-D staining is increased upon stimulation (IL-2 or anti-CD3/28)
• Intracellular flow staining results showing upregulation of protein expression have been confirmed by immunoblotting
• Furthermore, stimulated Perforin Knock-out (developed by Walsh) splenocytes do not stain with eBioOMAK-D nor is any protein detectable by western blotting with eBioOMAK-D as well as other anti-mouse perforin antibodies
• Please note that the Kagi perforin knock-out mice may synthesize a truncated form of the protein which may be recognized by eBioOMAK-D
• In IL-2 stimulated mouse splenocytes, NK cells (as determined by CD49b staining) contain perforin while CD8 cells contain little to none and can vary with culture conditions
• This has been confirmed by staining and western blotting the two populations using both OMAK-D and P1-8 antibodies
• Perforin is one of the major cytolytic proteins of cytolytic granules
• Perforin is a cytolytic mediator and is stored in and released by cytoplasmic granules
• Moreover, perforin is involved in immune defense against tumors and virus infections as mediated by cytotoxic lymphocytes
• Perforin is a 555 amino acid protein with a 21 amino acid signal peptide, and has a molecular weight of 70 to 75 kD
• Perforin is a pore forming protein with a mechanism of transmembrane channel formation similar to C9, and homology between perforin and C9 have been demonstrated
• Studies show that perforin is expressed only in killer cell lines and not in helper T lymphocytes or other tumor cells tested
• Perforin is known to be a key effector molecule for T-cell- and natural killer-cell-mediated cytolysis
• Defects in the perforin gene cause familial hemophagocytic lymphohistiocytosis type 2 (HPLH2), a rare and lethal autosomal recessive disorder of early childhood
• Alternative splicing results in multiple transcript variants of perforin.