Production and characterization of monoclonal antibodies raised against the membrane ecdysone receptor from the anterior silk gland of the silkworm, Bombyx mori
Journal Title: IOSR Journal of Pharmacy (IOSRPHR) - Year 2014, Vol 4, Issue 6
Abstract
Several evidences for nongenomic action of ecdysteroids through membrane receptor (mEcR) in programmed cell death (PCD) of the Bombyx mori anterior silk glands (ASGs) have been reported. The isolation, characterization and sequencing of mEcR still remain elusive. In the present work, monoclonal antibodies against mEcR has been raised, for the first time, and expected to facilitate any further molecular characterization of that novel receptor. The solubilized ASGs membrane fractions were used for immunizing mice. Three out of 4 injected mice showed immune-reactive signal, as revealed by dot blot assay. Out of the 3 mice, only one mouse sera showed to react with [3H]PonA binding protein, as revealed by double antibody capture assay (DACA). Out of 117 primary hybridoma cell lines prepared only 43 were selected by using dot blot assay, while 7 out of the 43 were selected by using DACA. From the 7 selected clones, clone (13-2) of the highest bound dpm in DACA was used for limiting dilution assay. The pattern of immune-reactivity of 7 out of 8 of the diluted cultures were very similar and showed two signal bands of 65 and 42 kDa, as revealed by western blot analysis. From which, a single clone designated (13-2-23) of the highest bound dpm in DACA was reacted with a single protein band of 42 kDa, suggesting its probable specificity to the extracellular binding domain of mEcR. This work will pave the way for more understanding of the molecular mechanism of ecdysteroid rapid actions during PCD of ASGs in Bombyx mori
Authors and Affiliations
Mohamed Elmogy , Azza M. Elgendy
Keywords
Related Articles
- EP ID EP121416
- DOI -
- Views 64
- Downloads 0
Attitude of Pharmacists to the Pharmaceutical care concept in rivers and bayelsa states of Southern Nigeria
Pharmaceutical care education was introduced into the country about a decade ago. There is a need to assess the impact of the education and information on the attitude of practicing pharmacists from time to time....
Candidate Molecules as Tumor Suppressor for Human Uterine Mesenchymal Tumor
Uterine leiomyosarcoma (Ut-LMS) develops more often in myometrium of the uterine body than in the uterine cervix. The development of gynecologic tumors is often correlated with female hormone secretion; however,...
Quality Of Life of Hypertensive Patients on Different Types of Antihypertensive Medications
: More than three quarter of the total death in the UAE is due to cardiovascular diseases. Health Related Quality of Life (HRQL) is a broad multidimensional concept that usually includes subjective evaluations...
In-vivo antipyretic activity of methanolic extracts of root and leaves of Morinda angustifolia Roxb.
ABSTRACT: Bangladesh is a country of vast natural resources and plants of utmost values. Over the years, natural products based medical practitioners have been using numerous plants as the panacea of wide rang...
Carotenoids From Red Fruit (Pandanus Conoideus Lam.) Extract Are Bioavailable : A Study In Rats
: Red Fruit (Pandanus conoideus Lam.) is a Papuan indigenous fruit. This fruit has the potential of becoming functional food because of its carotenoid compounds that acts as an antioxidant and provitamin A. The h...