Exbio — Research products — Recombinant allergens — Venom — rApi m 1 (Apis mellifera 1.0101)

rApi m 1 (Apis mellifera 1.0101)

Regulatory status

RUO

Format

Purified Azide Free

Application

ELISA, WB

Variant

0.1 mg

10-P195-C100

In stock

242.00 USD

Variant

0.1 mg

10-P195-C100

In stock

242.00 USD

Contact distributor

Product details

Images

SDS download

Application

ELISA, WB

Description

Recombinant protein rApi m 1 is expressed in S2 cell (Drosophila). DNA sequence encoding 148 AAs was fused with Strep-tag at the N-terminus. A calculated molecular mass of recombinant protein is 16,8 kDa.

Contents

Recombinant protein rApi m 1 - 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.

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Recombinant allergen rApi m 1 purity verification. 5 µg of rApi m 1 with > 95 % purity checked by Coomassie Brilliant Blue stained SDS-PAGE.

Reactivity of rApi m 1 with sIgE antibodies in plasma of a patient with confirmed presence (routine clinical test) of specific IgE antibodies to honey bee venom allergen extract.

Western blotting analysis was performed on recombinant allergen rApi m 1 mixed and heated (100°C, 5 min) with reducing SDS loading buffer. Sample was resolved using 15% SDS-PAGE gel. Nitrocellulose membrane blot was stained with Ponceau S to verify protein transfer. After thorough washing and blocking steps, the membrane was incubated with plasma of an allergic patient (1:10). Captured rApi m 1 sIgE antibodies were detected by biotinylated anti-human IgE secondary antibody (2 μg/ml) followed by IRDye 800CW Streptavidin (1:5000).