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Molecular cloning, stable expression and cellular localization of human α1-adrenergic receptor subtypes: effect of charcoal/dextran treated serum on expression and localization of α1D -adrenergic receptor

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Abstract

The cDNAs encoding for three subtypes of adrenergic receptors, α1A-, α1B- and α1D-ARs, were cloned and expressed in HEK 293 cells. Expression of α1A- and α1B-AR subtypes in HEK 293 cells was stable even with increased passages but that of α1D-AR was not. Cellular localization studies using immunofluorescence and flow cytometry revealed that expression of α1A- and α1B-ARs was primarily localized on the cell membrane whereas expression of α1D-AR was␣predominantly intracellular. Our studies clearly demonstrated that the culturing of the recombinant cell lines expressing α1D-AR in charcoal/dextran treated fetal bovine serum (FBS) resulted in targeting of α1D-AR to the cell membrane and thus, significantly improving its stability and availability for ligand binding studies.

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Acknowledgement

We thank Dr. Kedar Padmakar Purnapatre for critical reading of the manuscript.

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Authors and Affiliations

  1. Department of Biotechnology, Ranbaxy Research Laboratories, Plot No. 20, Sector 18, Udyog Vihar Industrial Area, Gurgaon, 122001, Haryana, India

    Sunil K. Khattar, Roop Singh Bora, Priyanka Priyadarsiny, Aarti Gautam, Dikshi Gupta & Kulvinder Singh Saini

  2. Department of Pharmacology, Ranbaxy Research Laboratories, Plot No. 20, Sector 18, Udyog Vihar Industrial Area, Gurgaon, 122001, Haryana, India

    Atul Tiwari, Kamna Nanda, Rahul Singh, Anita Chugh & Vinay Bansal

  3. New Drug Discovery Research, Ranbaxy Research Laboratories, Plot No. 20, Sector 18, Udyog Vihar Industrial Area, Gurgaon, 122001, Haryana, India

    Kasim Mookhtiar

Authors
  1. Sunil K. Khattar
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  2. Roop Singh Bora
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  3. Priyanka Priyadarsiny
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  8. Rahul Singh
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  9. Anita Chugh
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  10. Vinay Bansal
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  11. Kasim Mookhtiar
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  12. Kulvinder Singh Saini
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Corresponding author

Correspondence to Sunil K. Khattar.

Additional information

Sunil M. Khattar, Roop Singh Bora and Priyanka Priyadarsiny contributed equally to this work.

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Khattar, S.K., Bora, R.S., Priyadarsiny, P. et al. Molecular cloning, stable expression and cellular localization of human α1-adrenergic receptor subtypes: effect of charcoal/dextran treated serum on expression and localization of α1D -adrenergic receptor. Biotechnol Lett 28, 1731–1739 (2006). https://doi.org/10.1007/s10529-006-9148-x

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  • DOI: https://doi.org/10.1007/s10529-006-9148-x

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