Fel d 1 recombinant protein

Recombinant allergen Fel d 1 for Felis domesticus (cat)

Cat. Num. AAA14241

• Applications: ELISA
• Synonyms: Fel d 1; N/A; Recombinant allergen Fel d 1 for Felis domesticus (cat); Fel d 1 recombinant protein

• Host: Pichia pastoris
• Form/Format: Dry powder (stabilized with 5 % trhalose) ; 20 mM HEPES buffer pH 8 and 0.15 M NaCl (Before lyophilization)
• Concentration: 1.92 mg/ml (varies by lot)

• Applicable Applications for Fel d 1 recombinant protein: ELISA (EIA)
• Application Notes: Possible Applications: Western Blot (WB), Dot Blot (DB), Indirect ELISA, Positive Control in Direct ELISA, Chemiluminescence Immunoassays (CLIA), Lateral Flow (LF). Where this product has not been tested for use in a particular technique, this does not necessarily include its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates. The lab strongly recommends utilizing anti-CCD (cross-reactive-carbohydrate determinants) absorbent to eliminate the anti-CCD IgE antibodies present in normal human sera to prevent cross-reaction of this glycoprotein. By using this absorbent, reactions resulting from CCDs are stopped. In the specific case, you measure IgG with an allergen produced in P. pastoris, the lab recommends using an anti-CCD sorbent (Please Inquire) to avoid cross-reaction with antibodies in sera which react with fungal high mannose glycans.
• Batch Composition: - Components: his-Fel d 1; - Composition: recombinant allergen with a his-tag
• Absence of Precipitation After a Freezing and Thawing Cycle: OK
• Recombinant Allergen: Fel d 1 is the major allergen of Felis domesticus (cat) (Ohman et al., 1976).
• Observations: Proteins should be maintained frozen at high concentrations. In order to defrost the protein, maintain the aliquot at 25°C without shaking to avoid aggregation. Prior to making test dilutions and after defrosting the protein, it 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 0.461 ml of sterile double-distilled water for a final concentration of 1.92 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 glass-walls of the container. Upon reconstitution, leave the solution at least 15min. homogenizing with a mild agitation at 4°C. Avoid vigorous shaking that can cause foaming ans protein denaturation. With this reconstitution, the protein will be maintained at 8. It is recommended that the users carry out their absorbance determinaters to avoid concentrations variabilites due to the equipment used, mainly in reproducibility analysis.
• Preparation and Storage: Upon receipt , store at 4°C to -20°C in vertical position, avoiding all possible humidity and maintaining the viasl dry. Once reconstituted, it shold be aliquoted to avoid freezing and thawing cycles and stored at -20°C to -80°C.

SDS-PAGE

(Purity Control in SDS-PAGE: 15%SDS-PAGE analysis (15%) of 10 ul of recombinant allergen. Purity is >95% as determined by gel electrophoresis.)

ELISA

(Protein concentration determined espectrophotometricallyThis protein does not contain any Trp residues. Experience shows that this could result in more than 10% error in the computed extinction coefficient. Therefore, we have measured the protein concentration by using the colorimetric assay based on the interaction between Coomassie brilliant blue and the arginine and aromatic residues (Bradford Method) and its maximum absorption shifts from 470 nm to 595 nm. The standard curve was performed with the protein BSA.)

Related Product Information for Fel d 1 recombinant protein

The Felis domesticus uteroglobin Fel d 1 has been prepared as the recombinant mature protein composed of two chains and fused to a his-tag. Fel d 1 chain 1 and chain 2 are fused by a peptide linker y N-glycosylation motif at position 103 was mutated. Tetramer composed of covalently linked heterodimers of chains 1 and 2.

References

Ohman, J. L., Lowell, F. C., Bloch, K. J. and Kendall, S. Allergens of mammalian origin V. properties of extracts derived from domestic cat. 1976, Clin. Allergy 6:419-28.
Calculation of protein extinction coefficients from amino acid sequence data. Anal Biochem. 1989 Nov 1;182(2):319-26.