Perforin 1 (PRF1) Active Protein

Active Perforin 1 (PRF1)

Cat. Num. AAA2104734

• Applications: Cell Culture, Activity Assays
• Purity: >92%
• Synonyms: Perforin 1 (PRF1); Active Perforin 1 (PRF1); PRF1; Perforin 1 (PRF1) active protein

• Host: E Coli
• Purity/Purification: >92%

• Applicable Applications for Perforin 1 (PRF1) active protein: Cell Culture, Activity Assays
• Residues: Val40~Lys355
• Tag: Two N-terminal Tags, His-tag and GST-tag
• Buffer Formulation: 20mM Tris, 150mM NaCl, pH8.0, containing 0.05% sarcosyl and 5% trehalose.
• Predicted isoelectric point: 7.4
• Reconstitution: Reconstitute in 20mM Tris, 150mM NaCl (pH8.0) to a concentration of 0.1-1.0 mg/mL. Do not vortex.
• Activity: Perforin 1 (PRF1) is a pore forming cytolytic protein found in the granules of cytotoxic T lymphocytes (CTLs) and NK cells. Upon degranulation, perforin binds to the target cell's plasma membrane, and oligomerises in a Ca2+ dependent manner to form pores on the target cell. Perforin is thought to act by creating holes in the plasma membrane which triggers an influx of calcium and initiates membrane repair mechanisms. These repair mechanisms bring perforin and granzymes into early endosomes. Besides, Granzyme A (GZMA) has been identified as an interactor of PRF1, thus a binding ELISA assay was conducted to detect the interaction of recombinant mouse PRF1 and recombinant mouse GZMA. Briefly, PRF1 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100uL were then transferred to GZMA-coated microtiter wells and incubated for 2h at 37 degree C. Wells were washed with PBST and incubated for 1h with anti-PRF1 pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37 degree C. Finally, add 50uL stop solution to the wells and read at 450nm immediately. The binding activity of PRF1 and GZMA was shown in Figure 1, and this effect was in a dose dependent manner.
• Preparation and Storage: Store at 2-8 degree C for one month. Aliquot and store at -80 degree C for 12 months.

Activity Data

(Perforin 1 (PRF1) is a pore forming cytolytic protein found in the granules of cytotoxic T lymphocytes (CTLs) and NK cells. Upon degranulation, perforin binds to the target cell's plasma membrane, and oligomerises in a Ca2 dependent manner to form pores on the target cell. Perforin is thought to act by creating holes in the plasma membrane which triggers an influx of calcium and initiates membrane repair mechanisms. These repair mechanisms bring perforin and granzymes into early endosomes. Besides, Granzyme A (GZMA) has been identified as an interactor of PRF1, thus a binding ELISA assay was conducted to detect the interaction of recombinant mouse PRF1 and recombinant mouse GZMA. Briefly, PRF1 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100muL were then transferred to GZMA-coated microtiter wells and incubated for 2h at 37 degree C. Wells were washed with PBST and incubated for 1h with anti-PRF1 pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37 degree C. Finally, add 50uL stop solution to the wells and read at 450nm immediately. The binding activity of PRF1 and GZMA was shown in Figure 1, and this effect was in a dose dependent manner.Figure. The Binding Activity of PRF1 with GZMA.)