rPhl p 7 (Phleum pratense 7.0101) Antibody

SKU: orb1088532

Description

rPhl p 7 (Phleum pratense 7.0101) Antibody

Images & Validation

• Tested Applications: ELISA, FC, WB
• Application Notes:
  Recombinant protein rPhl p 7 is expressed in Escherichia coli. DNA sequence encoding 90 AAs was fused with Strep-tag at the N-terminus. A calculated molecular mass of recombinant protein is 10 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 rPhl p 7 purity verification. 5 µg of rPhl p 7 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 rPhl p 7 and allergen-specific human plasma IgG4 antibodies of Phleum pratense 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 rPhl p 7.

Flow cytometry dot-plot staining pattern of rPhl p 7 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 Phl p 5, Phl p 7, and Phl p 12 was analysed in plasma of a patient with confirmed presence (routine clinical test) of specific IgE antibodies to Timothy grass pollen allergen extract. Western blotting analysis was performed on recombinant allergens rPhl p 5, rPhl p 7, and rPhl p 12 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 Phl p 7.

Quick Database Links

UniProt

O82040

NCBI Gene ID

35