Pim proteins (Pim-1, Pim-2 and Pim-3) are oncogene-encoded serine/threonine kinases (1). Pim-1, a serine/threonine kinase highly expressed in hematopoietic cells, plays a critical role in the transduction of mitogenic signals and is rapidly induced by a variety of growth factors and cytokines (1-4). Pim-1 cooperates with c-Myc in lymphoid cell transformation and protects cells from growth factor withdrawal and genotoxic stress-induced apoptosis (5,6). Pim-1 also enhances the transcriptional activity of c-Myb through direct phosphorylation within the c-Myb DNA binding domain as well as phosphorylation of the transcriptional coactivator p100 (7,8). Hypermutations of the Pim-1 gene are found in B-cell diffuse large cell lymphomas (9). Phosphorylation of Pim-1 at Tyr218 by Etk occurs following IL-6 stimulation and correlates with an increase in Pim-1 activity (10). Various Pim substrates have been identified; Bad is phosphorylated by both Pim-1 and Pim-2 at Ser112 and this phosphorylation reverses Bad-induced cell apoptosis (11,12). The corresponding pim-1 gene encodes a pair of proteins through use of different translation initiation sites. Both larger 44 kDa (Pim-1L) and smaller 33 kDa (Pim-1S) proteins are active kinases, but differ in stability (13).
Intended Use:The Pim Kinase Antibody Sampler Kit provides an economical means to detect all three Pim kinases along with Bad and Phospho-Bad (Ser112). The kit contains enough primary and secondary antibody to perform two western blot experiments.
Specificity:The Pim-1 Rabbit mAb, Pim-2 Rabbit mAb, and Pim-3 Rabbit mAb detect endogenous levels of the respective proteins. The Pim antibodies do not crossreact with other Pim family members. Phospho-Bad (Ser112) Rabbit mAb detects endogenous levels of Bad only when phosphorylated at Ser112. It does not detect Bad phosphorylated at other sites, nor does it detect related family members. Bad Rabbit mAb detects endogenous levels of total Bad protein.
Kit Components:Kit (5x20ml)Pim-1 Mab (Rb x Hu): 1x20ulPim-2 Mab (Rb x Hu): 1x20ulPim-3 Mab (Rb x Hu): 1x20ulPhospho-Bad Mab (Ser112) (Rb x Hu): 1x20ulBad Mab (Rb x Hu): 1x20ulAnti-rabbit IgG, HRP-linked Antibody: 1x100ul
Applications:Suitable for use in Western Blot. Other applications not tested.
Recommended Dilution:Western Blot: 1:1000Optimal dilutions to be determined by the researcher.
Storage and Stability:Store liquid at -20°C. Store other components at 4°C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
