lipl32 recombinant protein

lipl32 recombinant

Cat. Num. AAA14238

• Applications: ELISA
• Synonyms: lipl32; N/A; lipl32 recombinant; RECOMBINANT ANTIGEN LipL32 from Leptospira interrogans; LipL32; lipl32 recombinant protein

• Host: Escherichia Coli.
• Form/Format: 20 mM phosphate buffer pH 6 and 0.15 M NaCl and 0.1 polyoxythylene (10) tridecyl ether.(before lyophilization)
• Concentration: 1.16 mg/ml (varies by lot)

• Applicable Applications for lipl32 recombinant protein: ELISA (EIA)
• Application Notes: Titration Curve By An ELISA Assay: The titer has been suggested in reference to an "in-house" ELISA kit performed over the first lot obtained. Each end user should carry out their own titration for their particular application.; ; Suggested Titer by ELISA: Up to 1:1,487, which corresponds to 0.78 ug/ml of protein concentration in plates for IgM detection.; Possible Applications: WB, DB, Indirect ELISA, positive control in direct ELISA, CLIA, lateral-flow. ; Where this product has not been tested for use in a particular technique, this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates.; ; Concentration Note: *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.
• Source: Escherichia coli
• Appearance: dry powder (stabilized with 15% trehalose)
• Quality Control: Protein Concentration Determined Espectrophotometrically: ; DO₂₈₀ = 1.08; A_{0.1 %} (=1 g/l) = 0.923
• Absence of precipitation after a freezing and thawing cycle: ok
• Observations: In some cases, purified proteins run at a molecular weight which is slightly different to the theoretically calculated molecular weight 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 experiment. In order to defrost the protein, maintain the aliquot at 25ºC without shaking to avoid aggregation. Prior 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.
• Reconstitution: With approx. 0.802 ml of sterile double-distilled water, a final concentration of approx. 1.16 mg/ml will be obtained. The solubilisation 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. Please keep in mind that the dinal volume of the protein solution may differ from the reconstitution 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 te certificate of analysis. 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 . 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 reconsitution , the protein will be maintained at pH6. It is recommended that the users carry out their absorbance determinations to avoid concentration variabilities due to the equiptment used,mainly in reproducibility analysis.
• Total Quantity per Aliquot: 1 mg
• Observations: 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 25ºC without shaking to avoid aggregation. Prior 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.
• Preparation and Storage: Protein is shipped at room temperature. Upon receipt, it should be stored at 4° to -20°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 -20ºC to -80ºC.

Application Data

(In this plot, the optical density at 450/620 nm for positive (blue) and negative (gray) IgM sera are compared for each concentration of the recombinant antigen. An appropriate statistical test of significance for the comparison of means between both groups, the Welch's test, is employed. Eligible concentrations for the use of the antigen should present statistically significant differences between positive and negative sera. This happens when the intervals at the top do not overlap and, equivalently, when the p-value at the bottom is below 0.05. In the present figure, all p-values are below 0.05 and thus the intervals do not overlap. Therefore, any of the showed concentrations can be used to distinguish between positive and negative sera.)

SDS-PAGE

(SDS-PAGE analysis (12%) of 1 ul of recombinant LipL32. Purity is >95% as determined by gel electrophoresis.)

Related Product Information for lipl32 recombinant protein

Description: The recombinant antigen LipL32 has been prepared as the mature antigen fused to a his-tag. It is produced from the ORF of the lipL32 gene which codifies the mayor outer membrane protein of the spirochete Leptospira interrogans without the leader peptide.

Product Categories/Family for lipl32 recombinant protein

BACTERIA

References

Haake, D., G. Chao, R. L. Zuerner, J. K. Barnett, D.Barnett, M. Mazel, J. Matsunaga, P. N. Levett, and C.A. Bolin. The leptospiral major outer membrane proteinLipL32 is a lipoprotein expressed during mammalianinfection. 2000, Infect Immun., 68(4):2276-2285.
Gill SC, von Hippel PH. Calculation of protein extinctioncoefficients from amino acid sequence data. AnalBiochem. 1989 Nov 1;182(2):319-26.

NCBI and Uniprot Product Information

• Molecular Weight: Determined by SDS-PAGE, the protein band is at the molecular marker of 35,000 Da, while relative molecular mass calculated from amino acid sequence is 34,961.4 Da.