Thermo Fisher Scientific Perforin Monoclonal Antibody (eBioOMAK-D), PE, eBioscience

Thermo Fisher Scientific Perforin Monoclonal Antibody (eBioOMAK-D), PE, eBioscience™

Applications

• Intracellular staining
• Flow cytometric analysis

Tested Dilution: 1 µg/test
Publications: View 47 publications

Product Specifications

Available Formats:

• Unconjugated
• APC
• FITC
• Request custom conjugation

Product Specific Information

The eBioOMAK-D antibody reacts with mouse perforin (pore-forming protein, pfp, Prf). Perforin is a cytolytic mediator found in cytotoxic T lymphocytes (CTL) and natural killer (NK) cells and plays a crucial role in immune response against tumors and viral infections.

By immunoblotting, eBioOMAK-D recognizes a ~70 kDa band in lysates of CTLL-2 mouse cytotoxic cell line and IL-2 stimulated mouse splenocytes. In multi-color intracellular flow cytometric analysis, staining intensity increases upon stimulation (IL-2 or anti-CD3/CD28). Knock-out splenocytes do not stain or show detectable protein, confirming specificity.

In IL-2 stimulated mouse splenocytes, NK cells (CD49b+) contain perforin, while CD8+ cells show variable expression depending on culture conditions. Anti-CD3/CD28 stimulation increases perforin expression in both NK and CD8 cells. eBioOMAK-D also cross-reacts with human perforin and co-stains CD56+ cells in PBMC.

Granzyme B expression does not always correlate with perforin; Granzyme B is expressed earlier and at higher levels. IL-2 stimulation increases Granzyme B expression >10,000-fold (mRNA) versus a 10–100-fold increase for perforin.

For optimal intracellular staining, use the Foxp3 buffer system (Product #00-5523). Other buffers may yield varying results.

Applications Reported: Intracellular staining followed by flow cytometric analysis.
Applications Tested: Tested on stimulated mouse splenocytes using the Foxp3/Transcription Factor Staining Buffer Set (Product #00-5523).

Recommended Protocol: BestProtocols® – Protocol B: One-step protocol for intracellular proteins.
Use ≤1 µg/test (100 µL final volume). Cell number: 10⁵–10⁸ cells/test. Titrate for optimal performance.

Excitation: 488–561 nm
Emission: 578 nm
Compatible Lasers: Blue, Green, Yellow-Green
Filtration: 0.2 µm post-manufacturing filtered

Target Information

Perforin is a major cytolytic protein stored in cytoplasmic granules of cytotoxic lymphocytes. It mediates immune defense against tumors and viral infections. The protein consists of 555 amino acids (including a 21 aa signal peptide) with a molecular weight of 70–75 kDa. Perforin forms transmembrane channels similar to complement component C9.

It is expressed only in killer cell lines, not in helper T or other tumor cells. Mutations in the perforin gene cause familial hemophagocytic lymphohistiocytosis type 2 (HPLH2), a rare autosomal recessive disorder. Multiple transcript variants exist due to alternative splicing.

For Research Use Only. Not for use in diagnostic procedures or resale without authorization.

제품 이미지