PMID: 8972729Nov 8, 1996Paper

Generation of antibodies specific for the RalA and RalB GTP-binding proteins and determination of their concentration and distribution in human platelets

Biochimica Et Biophysica Acta
O Jilkina, Rajinder P Bhullar

Abstract

Peptide specific polyclonal antibodies directed against C-termini of ras p21 related GTP-binding proteins, ralA and ralB, were generated. To assess antibody specificity, cDNAs coding for full length ralA and ralB were expressed in Escherichia coli as GST fusion proteins. Western blotting analysis using enhanced chemiluminescence technique confirmed that ralA and ralB antibodies were specific for their respective protein. To determine the concentration and distribution, varying amounts of GST-ralA and GST-ralB and, human platelet particulate and cytosolic proteins were loaded during Western blotting. The amount of ralA and ralB proteins in the platelet particulate fraction was determined to be 0.16 +/- 0.017 microgram/mg protein (n = 3) and 0.15 +/- 0.009 microgram/mg protein (n = 3) respectively. In the cytosol, only ralB protein was detected and its concentration was estimated to be 0.03 +/- 0.009 microgram/mg protein (n = 3). Both ralA and ralB proteins were isoprenylated in the presence of [3H] mevalonolactone plus rabbit reticulocyte lysate although radioactivity incorporated into ralA was three times higher than that associated with the ralB protein. Addition of geranylgeranyl pyrophosphate to the reaction mixture inhibite...Continue Reading

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Citations

Jan 16, 1999·Journal of Neuroscience Research·R P BhullarJ N Kanfer
Dec 14, 2001·Biochemical and Biophysical Research Communications·R S Sidhu, R P Bhullar
May 3, 2003·Biochemical and Biophysical Research Communications·Ranjinder S SidhuRajinder P Bhullar
Jul 14, 1999·Thrombosis Research·A McNicol, S J Israels
Apr 26, 2002·British Journal of Haematology·Qing ZhangRichard T Swank
Aug 30, 2005·Biochemical and Biophysical Research Communications·Ranjinder S SidhuRajinder P Bhullar
Sep 2, 2006·Biochimica Et Biophysica Acta·Olga Jilkina, Rajinder P Bhullar
Mar 11, 2008·Biochimica Et Biophysica Acta·Sherif M Elsaraj, Rajinder P Bhullar
Jul 7, 2007·Canadian Journal of Physiology and Pharmacology·Rajinder P BhullarOgnjen Grujic
Apr 9, 2005·The Journal of Biological Chemistry·Ranjinder S SidhuRajinder P Bhullar
May 30, 2002·The Journal of Biological Chemistry·Richard R CloughRajinder P Bhullar

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