rBet v 2 (Betula verrucosa 2.0101) Antibody

SKU: orb1088531

Description

rBet v 2 (Betula verrucosa 2.0101) Antibody

Research Area

Epigenetics

Images & Validation

• Tested Applications: ELISA, FC, WB
• Application Notes:
  Recombinant protein rBet v 2 is expressed in Escherichia coli. DNA sequence encoding 152 AAs was fused with Strep-tag at the N-terminus. A calculated molecular mass of recombinant protein is 16,2 kDa.

Key Properties

• Purification: Purified by sequential steps of ion exchange and affinity chromatography. Lyophilized from a 0.2 μm filtered solution in Storage buffer (100 mM Tris, 150 mM NaCl, 0.01% Tween 20, pH 8.0).
• Conjugation: Unconjugated

Storage & Handling

• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Expiration Date: 12 months from date of receipt.
• Disclaimer: For research use only

Recombinant allergen rBet v 2 purity verification. 5 µg of rBet v 2 with > 95 % purity checked by Coomassie Brilliant Blue stained SDS-PAGE.

ELISA (enzyme-linked immunosorbent assay) test was designed to prove the bond between the coated target recombinant allergen rBet v 2 and allergen-specific human plasma IgG4 antibodies of Betula verrucosa positive donor. A measurable signal was subsequently generated by the addition of biotin labeled anti-human IgG4 (detection) antibody, Streptavidin-HRP and substrate solution (TMB). The intensity of the signal is proportional to the amount of coated rBet v 2.

Flow cytometry dot-plot staining pattern of rBet v 2 recombinant allergen stimulated human peripheral whole blood lymphocytes and basophils of a proven allergic donor stained using anti-human CD63 (MEM-259) FITC and anti-human CD203c (NP4D6) PE antibodies of BasoFlowEx kit.

Presence of sIgE antibodies specific to recombinant allergens Bet v 1, Bet v 2, Bet v 4, Bet v 6, and Bet v 7 was analysed in plasma of a patient with confirmed presence (routine clinical test) of specific IgE antibodies to birch pollen allergen extract. Western blotting analysis was performed on recombinant allergens rBet v 1, rBet v 2, rBet v 4, rBet v 6, and rBet v 7 mixed and heated (100°C, 5 min) with reducing SDS loading buffer. Samples were resolved using 12% Tris-glycine gel electrophoresis. Nitrocellulose membrane blot was incubated with plasma of an allergic patient (1:10). Captured sIgE antibodies were detected by biotinylated anti-human IgE secondary antibody (2 μg/ml) followed by IRDye 800CW Streptavidin (1:5000). The analysed patient plasma contained sIgE antibodies specific to Bet v 2, Bet v 6, Bet v 7.

Quick Database Links

UniProt

P25816

NCBI Gene ID

35