| Products/Services Used | Details | Operation |
|---|---|---|
| DNA Sequencing | Following solubilization of the inclusion bodies using 8 M urea, the supernatant was loaded onto a Nickel column [GenScript (Nanjing) Co., Ltd.], washed with running buffer containing 20 mM Tris-HCl, 100 mM NaH2PO4, 10 mM imidazole and 8 M urea (pH 8.0), and eluted with elution buffer containing 20 mM Tris-HCl, 100 mM NaH2PO4, 250 mM imidazole and 8 M urea (pH 8.0). The nucleic acid sequence of mature Pla a 1 was synthesized by GenScript (Nanjing) Co., Ltd. (Nanjing, China) and was subcloned into a pET-28a vector (Novagen; Merck KGaA, Darmstadt, Germany) using EcoRI and XhoI sites, and the clone was verified by Sanger DNA sequencing, as described previously (21). | Get A Quote |
Platanus acerifolia pollen is considered an important source of airborne allergens in numerous cities. Pla a 1 is a major allergen from P. acerifolia pollen. The present study aimed to express and purify Pla a 1， and to prepare its monoclonal antibody. In the present study， the Pla a 1 gene was subcloned into a pET?28a vector and transformed into the ArcticExpress? (DE3) RP Escherichia coli host strain. The purified Pla a 1 was then used to immunize BALB/c mice. When serum detection was positive， spleen cells were isolated from the mice and fused with SP2/0 myeloma cells at a ratio of 10:1. Hybridoma cells were screened by indirect ELISA and limiting dilution. Positive cells were used to induce the fo... More
