LipL21 Recombinant Protein

Pichia pastoris

Dry powder (stabilized with 15% trehalose)

Upon receipt, it should be stored at 4°C to -20°C in vertical position, avoiding all possible humidity and maintaining dry. Once reconstituted, it should be aliquoted to avoid repeated freezing and thawing cycles and stored at -20°C to -80°C.

SDS-PAGE analysis determines that the protein band is between the molecular markers of 35,000 and 25,000 Da due to the glycosylation pattern, while relative molecular mass, calculated from amino acid sequence and without glycosylation, is 21,346.50 Da.

Leptospira interrogans lipL21 (Seenichamy et al., 2014)

Components: his-lipL21
Composition: recombinant antigen with a his-tag in its C-terminus

Components: Storage buffer before lyophilization
Composition: 20 mM phosphate buffer pH 8 and 0.1 M KCl

1. Protein concentration determined espectrophotometrically

DO₂₈₀ = 2.37
A _0.1% (=1 g/l) = 1.391
Concentration*: 1.70 mg/ml

* The measurement of the protein concentration has been performed with the theoretical extinction coefficient of the recombinant protein obtained from Gill and vonHippel, 1989. It is recommended that the users carry out their absorbance determinations to avoid equipment variabilities regarding final concentration, mainly in reproducibility analysis.

2. Absence of precipitation after a freezing and thawing cycle: ok

Reconstitute with a 0.51 ml of sterile double-distilled water for a final concentration of 1.70 mg/ml will be obtained. The solubilization of the cake should be developed for 15 min to allow a homogeneous protein solution, considering that part of the cake can be on the glass-walls of the container. Upon reconstitution, leave the solution at least 15 min homogenizing with a mild agitation at 4°C. Avoid vigorous shaking that can cause foaming and protein denaturation. With this reconstitution, the protein will be maintained at pH 8. It is recommended that the users carry out their absorbance determinations to avoid concentration variabilities due to the equipment used, mainly in reproducibility analysis

1 mg

Proteins should be maintained frozen at high concentrations. The dilution to be performed for ELISA assays should be made with a small quantity of protein, the same day of the experiment. In order to defrost the protein, maintain the aliquot at 25°C without shaking to avoid alterations in the immobilization of the biomolecule to the solid surface.

Manufactured in an ISO 9001:2015 Certified Laboratory.