| Products Included | No. | Volume | Applicaton | Dilution | Reactivity | Homology† |
|---|---|---|---|---|---|---|
| Primary Cocktail | 7847 | 100 µl | Immunofluorescence (Immunocytochemistry), Immunofluorescence (Paraffin)‡ | 1:100 | Human | Monkey |
| Detection Cocktail | 7843 | 100 µl | Immunofluorescence (Immunocytochemistry), Immunofluorescence (Paraffin)‡ | 1:100 | N/A | N/A |
†Species predicted to react based on 100% sequence homology.
‡Immunofluorescence (Paraffin) protocol recommended unmasking buffer: Citrate
| Kit Analytes | Detection Dye | Ex(max) (nm) | Em(max) (nm) |
|---|---|---|---|
| Phospho-Histone H3 (Ser10) | Alexa Fluor® 488 | 495 | 519 |
| Cleaved-PARP (Asp214) | Alexa Fluor® 647 | 650 | 665 |
| α-Tubulin | Alexa Fluor® 555 | 555 | 565 |
IF-P
#P68431, #P68363, #P09874
8350, 10376, 142
Product Information
Storage
Specificity / Sensitivity
Species Reactivity:
Human
Source / Purification
Monoclonal antibodies were produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser10 of human histone H3, a synthetic peptide corresponding to residues surrounding Asp213 of human PARP, or with full-length chicken α-tubulin purified from brain extracts.
Product Description
Background
Apoptosis is a regulated physiological process leading to cell death. Initiator caspases cleave and activate downstream effector caspases that in turn cleave cytoskeletal and nuclear proteins like PARP, α-fodrin, DFF, and lamin A, which induce apoptosis (1). Cleavage of the nuclear poly (ADP-ribose) polymerase PARP occurs between Asp214 and Gly215, which separates the PARP amino-terminal DNA binding domain (24 kDa) from the carboxy-terminal catalytic domain (89 kDa) (2,3). PARP helps to maintain cell viability by playing key roles in many cellular processes, including DNA replication, repair, and recombination. Cleavage of PARP facilitates cellular disassembly and serves as a marker of cells undergoing apoptosis (4). Cell proliferation can be measured by studying the phosphorylation state of histone H3 and microtubule assembly during mitosis. Histone H3 is one of four distinct core histone proteins found in the nucleosome. Phosphorylation of histone H3 at Ser10, Ser28, and Thr11 is tightly correlated with chromosome condensation during both mitosis and meiosis (5-7). Heterodimers composed of α-tubulin and β-tubulin form globular tubulin subunits common to all eukaryotic cells. Tubulin polymers known as microtubules are fundamental cytosolic fibers important in mediating cellular movement, including meiotic/mitotic chromosome alignment, cytoplasmic membrane vesicle transport, and nerve-cell axon migration (8).
- Nicholson, D.W. (1999) Cell Death Differ 6, 1028-42.
- Lazebnik, Y.A. et al. (1994) Nature 371, 346-7.
- Nicholson, D.W. et al. (1995) Nature 376, 37-43.
- Oliver, F.J. et al. (1998) J Biol Chem 273, 33533-9.
- Hendzel, M.J. et al. (1997) Chromosoma 106, 348-60.
- Goto, H. et al. (1999) J Biol Chem 274, 25543-9.
- Preuss, U. et al. (2003) Nucleic Acids Res 31, 878-85.
- Westermann, S. and Weber, K. (2003) Nat Rev Mol Cell Biol 4, 938-47.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Applications Key
IF-P: Immunofluorescence (Paraffin)
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
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