Cell Signaling Technology

Product Pathways - Lymphocyte Signaling

Pim Kinase Antibody Sampler Kit #9779

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
Pim-1 (C93F2) Rabbit mAb #3247 40 µl W H M (R) (Mk) (B) 34 Rabbit IgG
Pim-2 (D1D2) XP® Rabbit mAb #4730 40 µl W IP IHC-P IF-IC 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 23 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  B=Bovine
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

The Pim-1 (C93F2) Rabbit mAb, Pim-2 (D1D2) XP® 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.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Pim-1 (C93F2) Rabbit mAb #3247.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Pim-3 (D17C9) Rabbit mAb #4165.

Western Blotting

Western Blotting

Western blot analysis of extracts from K-562, Raji, and KARPAS-620 cell lines using Pim-2 (D1D2) XP® Rabbit mAb #4730.


Western Blotting

Western Blotting

Western blot analysis of extracts from COS cells, untreated or TPA-treated, using Phospho-Bad (Ser112) (40A9) Rabbit mAb #5284 (upper), or Bad Antibody #9292 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Bad (D24A9) Rabbit mAb #9239.

Description

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.

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.

Background

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).

  1. Mikkers, H. et al. (2004) Mol Cell Biol 24, 6104-15.
  2. Selten, G. et al. (1986) Cell 46, 603-11.
  3. Meeker, T.C. et al. (1987) J Cell Biochem 35, 105-12.
  4. Dautry, F. et al. (1988) J Biol Chem 263, 17615-20.
  5. Möröy, T. et al. (1993) Proc Natl Acad Sci USA 90, 10734-8.
  6. Lilly, M. and Kraft, A. (1997) Cancer Res 57, 5348-55.
  7. Leverson, J.D. et al. (1998) Mol Cell 2, 417-25.
  8. Winn, L.M. et al. (2003) Cell Cycle 2, 258-62.
  9. Pasqualucci, L. et al. (2001) Nature 412, 341-6.
  10. Kim, O. et al. (2004) Oncogene 23, 1838-44.
  11. Aho, T.L. et al. (2004) FEBS Lett 571, 43-9.
  12. Yan, B. et al. (2003) J Biol Chem 278, 45358-67.
  13. Saris, C.J. et al. (1991) EMBO J 10, 655-64.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Protocols

Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

Products