Exbio — Research products — Recombinant allergens — Pollen — rPhl p 7 (Phleum pratense 7.0101)

rPhl p 7 (Phleum pratense 7.0101)

Regulatory status

RUO

Format

Purified Azide Free

Application

ELISA, FC, WB

Variant

0.1 mg

10-P176-C100

In stock

242.00 USD

Variant

0.1 mg

10-P176-C100

In stock

242.00 USD

Contact distributor

Product details

Description

Images

SDS download

Application

ELISA, FC, WB

Description

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.

Contents

Recombinant protein rPhl p 7 - 1 vial (0.1 mg), lyophilized.
Reconstitute with sterile deionized water with or without azide.

Preparation

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).

Storage and handling

Store at -20°C to -80°C. Reconstitute in sterile deionized water. Use reconstituted product immediately or aliquot for further storage at -20°C to -80°C.

Unless indicated otherwise, all products are For Research Use Only and not for diagnostic use. In vivo diagnostic or therapeutic applications are strictly forbidden. Products shall not be used for resale or transfer to third parties either as a stand-alone product or as a manufacture component of another product without written consent of EXBIO Praha, a.s. EXBIO Praha, a.s. will not be held responsible for patent infringement or any other violations of intellectual property rights that may occur with the use of the products. Orders for all products are accepted subject to the Term and Conditions available at www.exbio.cz. EXBIO, EXBIO Logo, and all other trademarks are property of EXBIO Praha, a.s. © 2019 EXBIO Praha, a.s. All rights reserved.

UniProt ID O82040

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.

Recombinant allergen rPhl p 7 purity verification. 5 µg of rPhl p 7 with > 95 % purity checked by Coomassie Brilliant Blue stained SDS-PAGE.

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% SDS-PAGE gel. 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.