CFP10:ESAT6 Recombinant Protein

20 mM phosphate buffer pH 8 and 0.15 M NaCI

1.45 mg/mL (lot specific)

Protein is shipped at room temperature
Upon arrival, it should be stored at 4 degree to -20 degree C in vertical position, avoiding all possible humidity and maintaining the vials dry. Once reconstituted, it should be aliquoted to avoid repeated freezing and thawing cycles and stored at -20 degree C to -80 degree C.

Determined by SDS-PAGE, the protein band is between molecular markers of 25,000 and 18,400 Da, while relative molecular mass calculated from amino acid sequence is 22,100.0 Da.

Escherichia coli

1mg

M.tuberculosis chimeric antigen CFP10:ESAT6 (Bekmurzayevaet al., 2013)

Components:
his-CFP10:ESAT6
Composition:
Recombinant chimeric antigen with a his-tag in its N-terminus.

Ok

Protein concentration determined Espectrophotometrically:
Do₂₈₀= 1.64
A_0.1%(1 g/l)=1.13

Dry powder (stabilized with 15% trehalose)

In some cases, purified proteins run at a molecular weight which is slighty different to the theoretically calculated molecular wrigfht maybe due to the his-tag presence, which can produce a delay in SDS-PAGE. 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 expirament. In order to defrost the protein, maintain the aliquot at 25 degree C without shaking to avoid aggregation. Prior to making test dilutions and after defrosting the protein, is recommended to remove possible protein aggregates by centrifuging the stock solution, avoiding alterations in the immobilization of the biomolecule to the solid surface.

With 0.630 ml of sterile double-distilled water, a final concentratio of 1.45 mg/mL will be obtained. The solubilization of the cake should be developed for 15 minutes to allow a homogenous protein solution, considering that part of the cake can be on the galss-walls of the container. Please keep in mind that the final volume of the reconstituted protein solution may differ from the reconstituition volume mentioned in the instructions due to the hygroscopic nature of trehalose. As the protein is reconstituted, the final volume may slightly increase to reach the specified amount mentioned in the certificate of analysis. Upon reconstitution, leave the solution at least 15min. homogenizing with a mild agitation at 4°C. Avoid vigourous shaking that can cause foaming and protein denaturation. After those minutes, centrifuge the vial to ensure that all the product remains at the base and do not lose any of it on the walls. With this reconstituion, the protein will be maintained at pH 8. It is recommended that the users carry us out their absorbance determinations to avoid concentration varibabilities due to the equipment used, mainly in reproducibility analysis.

Manufactured in an ISO 9001:2015 Certified Laboratory.