Revision 1

#9770Store 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
c-IAP1 (D5G9) Rabbit mAb 7065 20 µl 62 kDa Rabbit IgG
c-IAP2 (58C7) Rabbit mAb 3130 20 µl 70 kDa Rabbit IgG
Survivin (71G4B7) Rabbit mAb 2808 20 µl 16 kDa Rabbit IgG
XIAP (3B6) Rabbit mAb 2045 20 µl 53 kDa Rabbit IgG
Livin (D61D1) XP® Rabbit mAb 5471 20 µl 34, 36 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 IAP Family Antibody Sampler Kit provides an economical means to investigate the expression of various IAP family members within the cell. The kit contains enough primary and secondary antibodies 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

The inhibitor of apoptosis protein (IAP) family consists of an evolutionarily conserved group of apoptosis inhibitors containing a conserved 70 amino acid BIR (baculovirus inhibitor repeat) domain (1,2). Human members of this family include c-IAP1, c-IAP2, XIAP, survivin, livin, and NAIP. Overexpression of IAP family members, particularly survivin and livin, in cancer cell lines and primary tumors suggests an important role for these proteins in cancer progression (3-5). In general, the IAP proteins function through direct interactions to inhibit the activity of several caspases, including caspase-3, caspase-7, and caspase-9 (5,6). In addition, binding of IAP family members to the mitochondrial protein Smac blocks their interaction with caspase-9, thereby allowing the processing and activation of the caspase (2).

  1. Deveraux, Q.L. and Reed, J.C. (1999) Genes Dev 13, 239-52.
  2. Deveraux, Q.L. et al. (1998) EMBO J 17, 2215-23.
  3. Altieri, D.C. et al. (1999) Lab Invest 79, 1327-33.
  4. Tamm, I. et al. (2000) Clin Cancer Res 6, 1796-803.
  5. Kasof, G.M. and Gomes, B.C. (2001) J Biol Chem 276, 3238-46.
  6. Deveraux, Q.L. et al. (1997) Nature 388, 300-4.

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.

    Limited Uses

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    Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

    Revision 1
    #9770

    IAP Family Antibody Sampler Kit

    IAP Family Antibody Sampler Kit: Image 1 Expand Image
    Simple Western™ analysis of lysates (1.0 mg/mL) from Jurkat cells using Survivin (71G4B7) Rabbit mAb #2808. The virtual lane view (left) shows the target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (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.
    IAP Family Antibody Sampler Kit: Image 2 Expand Image
    Simple Western™ analysis of lysates (1.0 mg/mL) from Ramos cells using c-IAP2 (58C7) Rabbit mAb #3130. The virtual lane view (left) shows a single target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (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.
    IAP Family Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of extracts from HeLa, Raji and COS cell lines, using XIAP (3B6) Rabbit mAb.
    IAP Family Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of lysates from Raji (human), BaF3 (mouse) and C6 (rat) cell lines using Survivin (71G4B7E) Rabbit mAb.
    IAP Family Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of extracts from Raji, Ramos and SR cell lines, using c-IAP2 (58C7) Rabbit mAb.
    IAP Family Antibody Sampler Kit: Image 6 Expand Image
    Western blot analysis of extracts from various cell lines using Livin (D61D1) XP® Rabbit mAb.
    IAP Family Antibody Sampler Kit: Image 7 Expand Image
    Western blot analysis of extracts from various cell lines using c-IAP1 (D5G9) Rabbit mAb.
    IAP Family Antibody Sampler Kit: Image 8 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.
    IAP Family Antibody Sampler Kit: Image 9 Expand Image
    Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® Survivin siRNA II (+), using Survivin (71G4B7E) Rabbit mAb #2808 and α-Tubulin (11H10) Rabbit mAb #2125. Survivin (71G4B7E) Rabbit mAb confirms silencing of survivin expression and α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of survivin siRNA.
    IAP Family Antibody Sampler Kit: Image 10 Expand Image
    Confocal immunofluorescent analysis of SK-MEL-28 cells (left) and HeLa cells (right) using Livin (D61D1) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
    IAP Family Antibody Sampler Kit: Image 11 Expand Image
    Western blot analysis of extracts from HeLa cells, mock transfected (-) or transfected with human c-IAP1 (+), using c-IAP1 (D5G9) Rabbit mAb.
    IAP Family Antibody Sampler Kit: Image 12 Expand Image
    Immunohistochemical analysis of paraffin-embedded human transitional epithelial carcinoma of the bladder using Survivin (71G4B7E) Rabbit mAb.
    IAP Family Antibody Sampler Kit: Image 13 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lung carcinoma showing nuclear localization using Survivin (71G4B7E) Rabbit mAb #2808.
    IAP Family Antibody Sampler Kit: Image 14 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Survivin (71G4B7E) Rabbit mAb in the presence of control peptide (left) or Survivin Blocking Peptide #1037 (right).
    IAP Family Antibody Sampler Kit: Image 15 Expand Image
    Immunohistochemical analysis of paraffin-embedded human pituitary adenoma using Survivin (71G4B7E) Rabbit mAb.
    IAP Family Antibody Sampler Kit: Image 16 Expand Image
    Confocal immunofluorescent analysis of HeLa cells using Survivin (71G4B7E) Rabbit mAb (green). Mitochondria have been labeled with MitoTracker® Red CMXRos (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    IAP Family Antibody Sampler Kit: Image 17 Expand Image
    Flow cytometric analysis of untreated Jurkat cells using Survivin (71G4B7E) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).