Abstract
Glucocorticoids (GC), steroid hormones, induce apoptosis and cell cycle arrest in lymphoid tissues and
hence widely employed in the therapy of children with acute lymphoblastic leukaemia (ALL). We reported a proapoptotic novel BH3-containing transcript, termed Bimbam, from children with ALL and cell lines. Bimbam is
similar to Bim in molecular mass and co-migrates on SDS-PAGE as recognized by anti Bim antibody. The
current study was carried out to separate the both isoforms and whether Bimbam exist at endogenous level. To
separate both splice variant, Bimbam subclone#4 and Bim subclone#17 were resolved on long SDS-PAGE and
the western blot was recognized by anti Bim antibody but it could not serve the purpose. Further, we applied 2D
Gel Electrophoresis to discriminate Bimbam and Bim. This technique includes; Iso-electric point that resolves the
protein on the basis of charges of amino acid in one direction and molecular mass in the other direction. When
whole cell proteins from Bim subclone #17, Bimbam subclone # 4 and their mixture were probed with anti Bim
antibody on 2D blot, the protein spot corresponding to both proteins could be recognized. To confirm the
endogenous existence of Bimbam, dexamethasone treated Pre B cells were subjected to 2D Gel electrophoresis
and the blot was subsequently probed with anti Bim antibody. The presence of double spots at the level of 25kd
provides convincing evidence regarding the presence of endogenous Bimbam but the conclusive evidence can
only be obtained through knowk down by Bimbam siRNA
MUHAMMAD MANSHA, MUHAMMAD TARIQ, MUHAMMAD WASIM. (2016) Identification of Endogenous “Bimbamˮ A novel GC-induced BH3-Domain containing Isoform through 2D Gel Electrophoresis, Biologia – Journal of Biological Society of Pakistan, Volume 62 (I), Issue 1.
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