Revision 1

#9779Store at -20C

1 Kit

(5 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
Pim-1 (C93F2) Rabbit mAb 3247 20 µl 34 kDa Rabbit IgG
Pim-2 (D1D2) Rabbit mAb 4730 20 µl 40, 38, 34 kDa Rabbit 
Pim-3 (D17C9) Rabbit mAb 4165 20 µl 35 kDa Rabbit IgG
Phospho-Bad (Ser112) (40A9) Rabbit mAb 5284 20 µl 23 kDa Rabbit IgG
Bad (D24A9) Rabbit mAb 9239 20 µl 23 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

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 two western blot experiments.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

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.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    使用に関する制限

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    Revision 1
    #9779

    Pim Kinase Antibody Sampler Kit

    Pim Kinase Antibody Sampler Kit: Image 1 Expand Image
    Simple Western™ analysis of lysates (0.1 mg/mL) from serum-starved COS cells treated with TPA (200 nM, 20 min) using Phospho-Bad (Ser112) (40A9) Rabbit mAb #5284. The virtual lane view (left) shows a single target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
    Pim Kinase Antibody Sampler Kit: Image 2 Expand Image
    Simple Western™ analysis of lysates (1 mg/mL) from OVCAR8 cells using Bad (D24A9) Rabbit mAb #9239. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
    Pim Kinase Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of extracts from various cell lines using Pim-1 (C93F2) Rabbit mAb.
    Pim Kinase Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of extracts from various cell lines using Pim-3 (D17C9) Rabbit mAb.
    Pim Kinase Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of extracts from K-562, Raji and KARPAS-620 cell lines using Pim-2 (D1D2) Rabbit mAb.
    Pim Kinase Antibody Sampler Kit: Image 6 Expand Image
    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).
    Pim Kinase Antibody Sampler Kit: Image 7 Expand Image
    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.
    Pim Kinase Antibody Sampler Kit: Image 8 Expand Image
    Western blot analysis of extracts from various cell lines using Bad (D24A9) Rabbit mAb.
    Pim Kinase Antibody Sampler Kit: Image 9 Expand Image
    Western blot analysis of recombinant Pim-1, -2, and -3 using Pim-1 (C93F2) Rabbit mAb.
    Pim Kinase Antibody Sampler Kit: Image 10 Expand Image
    Western blot analysis of recombinant Pim-1, 2, and 3 using Pim-3 (D17C9) Rabbit mAb.
    Pim Kinase Antibody Sampler Kit: Image 11 Expand Image
    Western blot analysis of recombinant Pim-1, Pim-2 and Pim-3 kinases using Pim-2 (D1D2) Rabbit mAb.
    Pim Kinase Antibody Sampler Kit: Image 12 Expand Image
    Immunohistochemical analysis of paraffin-embedded human breast carcinoma, untreated (left) or lambda phosphatase treated (right), using Phospho-Bad (Ser 112) (40A9) Rabbit mAb.
    Pim Kinase Antibody Sampler Kit: Image 13 Expand Image
    Western blot analysis of extracts from COS-7 cells, untransfected () or transfected with a mouse Pim-3 construct (+), using Pim-3 (D17C9) Rabbit mAb.
    Pim Kinase Antibody Sampler Kit: Image 14 Expand Image
    Immunohistochemical analysis of paraffin embedded COS cells untreated (left) or TPA-treated (right), showing induced cytoplasmic staining using Phospho-Bad (Ser112) (40A9) Rabbit mAb.
    Pim Kinase Antibody Sampler Kit: Image 15 Expand Image
    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).
    Pim Kinase Antibody Sampler Kit: Image 16 Expand Image
    Immunohistochemical analysis of paraffin-embedded Non-Hodgkin's lymphoma, using Phospho-Bad (Ser112) (40A9) Rabbit mAb.
    Pim Kinase Antibody Sampler Kit: Image 17 Expand Image
    Flow cytometric analysis of COS cells, untreated (blue) or TPA/Calyculin A treated (green), using Phospho-Bad (Ser112) (40A9) Rabbit mAb.