Human Perforin-1 (PRF1) Protein

Proteins and Peptides

Perforin-1 (PRF1)

E. coli

Activity: Active
Biological Activity: A binding ELISA assay was carried out to detect the interaction of recombinant Human PRF1 and recombinant Human Calreticulin (CRT). PRF1 was serially diluted in PBS with 0.01% BSA, pH 7.4. 100 µl duplicates were then transferred to CRT-coated microplate wells and incubated for 2 h at 37°C. Wells were washed with PBST and incubated for 1 h with a polyclonal PRF1 antibody, then aspirated and washed 3 times. They were then incubated with a HRP-conjugated secondary antibody and aspirated/washed 3 times. TMB substrate solution was added to the wells, which were then incubated for 15-25 minutes at 37°C. 50 µl stop solution was added to the wells and the absorbance was measured at 450 nm. The binding activity of PRF1 and CRT was determined to be dose-dependent.
The activity of recombinant Human PRF1 Protein was also observed with a hemolysis assay measuring lysis of rabbit erythrocytes (RaE), compared with a negative control. Recombinant Human PRF1 was diluted 2-fold in 0.9% NaCl. 50 µl diluted PRF1 (12.5 µg/ml) and 10 µl 0.1 M CaCl2 were added to each well, followed by 50 µl 0.25% RaE. They were mixed gently and the plate was incubated for 20 h at 37°C, 5% CO2. For the negative control, 10 µl 0.9% NaCl was used instead of 10 µl 0.1 M CaCl2.
The minimal effective concentration of PRF1 was determined to be 12.5 μg/ml.
Sequence Fragment: Lys32-Phe316
Tag: N-terminal His tag and GST tag

Human

Unconjugated

Calculated MW: 61.5 kDa  Observed MW (SDS-PAGE): 62 kDa

Recombinant

> 80%

50 µg

100 µg

200 µg

500 µg

1 mg

Lyophilized

WB, SDS-PAGE

Prior to lyophilization: 20 mM Tris, 150 mM NaCl, pH 8.0, containing 0.01% Sarcosyl and 5% Trehalose.

Shipped within 5-7 working days.

Store lyophilized form at 2-8°C for up to 1 month. For longer periods, store lyophilized or liquid at -80°C. Avoid repeated freeze–thaw cycles.

No

P14222

Protein PRF1

RUO

THIS PRODUCT IS FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC, THERAPEUTIC OR COSMETIC PROCEDURES. NOT FOR HUMAN OR ANIMAL CONSUMPTION.
To keep the original salt concentration, we recommend reconstituting to the original concentration prior to lyophilization (see Concentration) in ddH2O. If a lower concentration is required, dilute in 20 mM Tris, 150 mM NaCl, pH 8.0. If a higher concentration is required, the product can be reconstituted directly in 20 mM Tris, 150 mM NaCl, pH 8.0, though please note that this will change the overall salt concentration. The stock concentration should be between 0.1-1.0 mg/ml. Do not vortex.
Endotoxin Level: < 1.0 EU per 1 µg (LAL method)
Concentration: Prior to lyophilization: 150 µg/ml