Cell Signaling Technology

Product Pathways - Apoptosis

SignalStain® Proliferation/Apoptosis IHC Sampler Kit #8109

Kit Includes Quantity Antigen Retrieval / Diluent Isotype
Cleaved Caspase-3 (Asp175) Antibody #9661 40 µl Citrate / SignalStain®
Antibody Diluent #8112		 Rabbit IgG
Phospho-Histone H3 (Ser10) Antibody #9701 40 µl Citrate / SignalStain®
Antibody Diluent #8112		 Rabbit IgG
PCNA (PC10) Mouse mAb #2586 40 µl Citrate / SignalStain®
Antibody Diluent #8112		 Mouse IgG2a
Survivin (71G4B7E) Rabbit mAb #2808 40 µl Citrate / SignalStain®
Antibody Diluent #8112		 Rabbit IgG
*SignalStain® Antibody Diluent #8112 25 ml
†SignalSlide™ Cleaved Caspase-3 (Asp175)
IHC Controls #8104		 1 Pack

*SignalStain® Antibody Diluent is supplied as a working solution and should be stored at 4ºC (packaged separately).
†Control slides should be stored at 4ºC (packaged separately).

Selected rabbit monoclonal antibodies produced under license (granting certain rights including those under U. S. Patent No. 5,675,063 and U.S.S.N. 11/476,277) from Epitomics, Inc.

Custom Ordering Details

The control slides that accompany this kit are cut freshly upon ordering. Domestic: Please allow up to three business days for your order to be processed and shipped. International: Please allow up to one week for your order to be processed and shipped.

Description

The SignalStain® Proliferation/Apoptosis IHC Sampler Kit from Cell Signaling Technology allows the researcher to examine paraffin-embedded tissues or cells with antibodies that will detect cellular apoptosis or proliferation. Each antibody is validated for use in immunohistochemical assays using multiple approaches. Also included in the kit are control slides that can be used to verify the performance of each antibody and a primary antibody diluent. Please see table above for recommended diluent for each antibody in this kit.

Specificity / Sensitivity

Each antibody in the SignalStain® Proliferation/Apoptosis IHC Sampler Kit detects endogenous levels of its target protein and does not cross-react with any related proteins.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a recombinant Protein A-PCNA fusion protein, or with a synthetic peptide corresponding to residues surrounding Cys60 of human Survivin. Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser10 of human histone H3, or with a synthetic peptide surrounding amino-terminal residues adjacent to Asp175 of human caspase-3. Antibodies are purified by protein A and peptide affinity chromatography.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin embedded Jurkat cell pellets, untreated (left) or etoposide-treated (right), using Phospho-Histone H3 (Ser10) Antibody, PCNA (PC10) Mouse mAb, Cleaved Caspase-3 (Asp175) Antibody and Survivin (71G4B7E) Rabbit mAb. Cell pellets are provided in the SignalSlide® Cleaved Caspase-3 (Asp175) IHC Controls.

Schematic

Schematic

Background

Caspase-3 (CPP-32, Apoptain, Yama, SCA-1) is a critical executioner of apoptosis, as it is either partially or totally responsible for the proteolytic cleavage of many key proteins such as the nuclear enzyme poly (ADP-ribose) polymerase (PARP) (1). Activation of caspase-3 requires proteolytic processing of its inactive zymogen into activated p17 and p12 fragments (2). Survivin is a 16 kDa anti-apoptotic protein highly expressed during fetal development and cancer cell malignancy (3). Survivin binds and inhibits caspase-3, controlling the checkpoint in the G2/M-phase of the cell cycle through inhibiting apoptosis and promoting cell division (4,5). Proliferating cell nuclear antigen (PCNA) is a member of the DNA sliding clamp family of proteins that assist in DNA replication (6). Multiple proteins involved in DNA replication, DNA repair, and cell cycle control bind to PCNA rather than directly associating with DNA, thus facilitating fast processing of DNA (reviewed in 7). PCNA protein expression is a well-accepted marker of proliferation, and PCNA (PC10) Mouse mAb has been used to assess PCNA levels in hundreds of scientific studies. Histone H3, H2A, H2B, and H4 make up the core histones of the nucleosome. In response to various stimuli, the amino-terminal tails of core histones undergo various post-translational modifications, including acetylation, phosphorylation, methylation and ubiquitination, which affect the accessibility of chromatin to transcription factors (8). Phosphorylation of Histone H3 at Ser10, Ser28 and Thr11 of histone H3 is tightly correlated with chromosome condensation during both mitosis and meiosis (9-11).

  1. Fernandes-Alnemri, T. et al. (1994) J Biol Chem 269, 30761-4.
  2. Nicholson, D.W. et al. (1995) Nature 376, 37-43.
  3. Reed, J.C. and Reed, S.I. (1999) Nat Cell Biol 1, E199-200.
  4. Li, F. et al. (1998) Nature 396, 580-4.
  5. Li, F. et al. (1999) Nat Cell Biol 1, 461-6.
  6. Kelman, Z. and O'Donnell, M. (1995) Nucleic Acids Res 23, 3613-20.
  7. Maga, G. and Hubscher, U. (2003) J Cell Sci 116, 3051-60.
  8. Jaskelioff, M. and Peterson, C.L. (2003) Nat Cell Biol 5, 395-9.
  9. Hendzel, M.J. et al. (1997) Chromosoma 106, 348-60.
  10. Goto, H. et al. (1999) J Biol Chem 274, 25543-9.
  11. Preuss, U. et al. (2003) Nucleic Acids Res 31, 878-85.

Application References

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Protocols

* Product-specific protocol.

Companion Products

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

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