Product Pathways - Lymphocyte Signaling
Pim Kinase Antibody Sampler Kit #9779
|9779S||1 Kit (5 x 40 µl)||---||In Stock||---|
|Kit Includes||Quantity||Applications||Reactivity||Homology†||MW (kDa)||Isotype|
|Pim-1 (C93F2) Rabbit mAb #3247||40 µl||W||H, M||R, Mk, B||34||Rabbit IgG|
|Pim-2 (D1D2) Rabbit mAb #4730||40 µl||W, IP||H||40, 38, 34||Rabbit|
|Pim-3 (D17C9) Rabbit mAb #4165||40 µl||W||H, M, R||Mk||35||Rabbit IgG|
|Phospho-Bad (Ser112) (40A9) Rabbit mAb #5284||40 µl||W, IHC-P, F||H, M, R, Mk||23||Rabbit IgG|
|Bad (D24A9) Rabbit mAb #9239||40 µl||W||H, M, R, Mk||B||23||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody #7074||100 µl||W||Goat|
†Species predicted to react based on 100% sequence homology.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), F=Flow Cytometry
Reactivity Key: H=Human, M=Mouse, R=Rat, Mk=Monkey
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Western blot analysis of extracts from COS cells, untreated or TPA-treated, using Phospho-Bad (Ser112) (40A9) Rabbit mAb (upper) or Bad Antibody #9292 (lower).
Western blot analysis of extracts from K-562, Raji and KARPAS-620 cell lines using Pim-2 (D1D2) Rabbit mAb.
Western blot analysis of extracts from various cell lines using Pim-1 (C93F2) Rabbit mAb.
Western blot analysis of extracts from various cell lines using Bad (D24A9) Rabbit mAb.
Western blot analysis of extracts from various cell lines using Pim-3 (D17C9) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, untreated (left) or lambda phosphatase treated (right), using Phospho-Bad (Ser 112) (40A9) Rabbit mAb.
Western blot analysis of recombinant Pim-1, Pim-2 and Pim-3 kinases using Pim-2 (D1D2) Rabbit mAb.
Western blot analysis of recombinant Pim-1, -2, and -3 using Pim-1 (C93F2) Rabbit mAb.
Western blot analysis of recombinant Pim-1, 2, and 3 using Pim-3 (D17C9) Rabbit mAb.
Immunohistochemical analysis of paraffin embedded COS cells untreated (left) or TPA-treated (right), showing induced cytoplasmic staining using Phospho-Bad (Ser112) (40A9) Rabbit mAb.
Western blot analysis of extracts from COS-7 cells, untransfected () or transfected with a mouse Pim-3 construct (+), using Pim-3 (D17C9) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma, using Phospho-Bad (Ser 112) (40A9) Rabbit Monoclonal Antibody preincubated with control peptide (left) or Phospho-Bad (Ser 112) Blocking Peptide (IHC Specific) #1026 (right).
Immunohistochemical analysis of paraffin-embedded Non-Hodgkin's lymphoma, using Phospho-Bad (Ser112) (40A9) Rabbit mAb.
Flow cytometric analysis of COS cells, untreated (blue) or TPA/Calyculin A treated (green), using Phospho-Bad (Ser112) (40A9) Rabbit mAb.
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 four western blot experiments.
Specificity / Sensitivity
The Pim-1 (C93F2) Rabbit mAb, Pim-2 (D1D2) Rabbit mAb, and Pim-3 (D17C9) Rabbit mAb detect endogenous levels of the respective proteins. The Pim antibodies do not crossreact with other Pim family members. Phospho-Bad (Ser112) (40A9) 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 (D24A9) Rabbit mAb detects endogenous levels of total Bad protein.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Val160 of human Pim-1, Cys266 of human Pim-2, Ser275 of human Pim-3, Ser112 of mouse Bad, and Pro102 of human Bad, respectively.
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).
- Mikkers, H. et al. (2004) Mol Cell Biol 24, 6104-15.
- Selten, G. et al. (1986) Cell 46, 603-11.
- Meeker, T.C. et al. (1987) J Cell Biochem 35, 105-12.
- Dautry, F. et al. (1988) J Biol Chem 263, 17615-20.
- Möröy, T. et al. (1993) Proc Natl Acad Sci USA 90, 10734-8.
- Lilly, M. and Kraft, A. (1997) Cancer Res 57, 5348-55.
- Leverson, J.D. et al. (1998) Mol Cell 2, 417-25.
- Winn, L.M. et al. (2003) Cell Cycle 2, 258-62.
- Pasqualucci, L. et al. (2001) Nature 412, 341-6.
- Kim, O. et al. (2004) Oncogene 23, 1838-44.
- Aho, T.L. et al. (2004) FEBS Lett 571, 43-9.
- Yan, B. et al. (2003) J Biol Chem 278, 45358-67.
- Saris, C.J. et al. (1991) EMBO J 10, 655-64.
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
U.S. Patent No. 5,675,063.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.