Products Included | No. | Volume | Applicaton | Dilution | Reactivity |
---|---|---|---|---|---|
Primary Cocktail | 8998 | 100 µl | Immunofluorescence (Immunocytochemistry) | 1:100 | Human Mouse Rat Monkey |
Detection Cocktail | 8997 | 100 µl | Immunofluorescence (Immunocytochemistry) | 1:100 | N/A |
Kit Analytes | Detection Dye | Ex(max) (nm) | Em(max) (nm) |
---|---|---|---|
Phospho-Akt (Ser473) | Alexa Fluor® 555 | 555 | 565 |
Phospho-p44/42 Erk1/2 (Thr202/Tyr204) | Alexa Fluor® 488 | 495 | 519 |
Phospho-S6 Ribosomal Protein (Ser235/236) | Alexa Fluor® 647 | 650 | 665 |
IF-IC
#P27361, #P31751, #Q9Y243, #P62753, #P28482, #P31749
5595, 208, 10000, 6194, 5594, 207
Product Information
Storage
Specificity / Sensitivity
Species Reactivity:
Human, Mouse, Rat, Monkey
Source / Purification
Monoclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Ser473 of human Akt, Thr202/Tyr204 of human p44 MAP kinase, and Ser235/Ser236 of human ribosomal protein S6.
Product Description
MW (kDa) | n/a |
Background
Akt, also referred to as PKB or Rac, plays a critical role in controlling the balance between survival and apoptosis (1-3). This protein kinase is a downstream effector of phosphoinositide-3 kinase (PI3K), and is activated by phospholipid binding and activation loop phosphorylation at Thr308 by PDK1 (4), as well as by phosphorylation within the carboxy terminus at Ser473 by the mTOR-rictor complex (TORC2) (5). This pathway is down-regulated following dephosphorylation of phosphatidyl-inositol 3,4,5 triphosphate by PTEN, as well as by deactivation of PI3K with targeted small molecule inhibitors such as wortmannin and LY294002 (2,3,6,7).
p70 S6 kinase, a mitogen activated Ser/Thr protein kinase downstream of PI3K and the mTOR-raptor complex (mTORC1), phosphorylates the S6 protein of the 40S ribosomal subunit leading to an increase in translation of mRNA transcripts that contain an oligopyrimidine tract in their 5’ untranslated region (8). These particular mRNA transcripts (5’TOP) encode proteins involved in cell cycle progression, as well as ribosomal proteins and elongation factors necessary for translation (8,9). Important S6 ribosomal protein phosphorylation sites include several residues (Ser235, Ser236, Ser240 and Ser244) located within a small, carboxy-terminal region of the S6 protein (10,11).
Both p44 and p42 mitogen-activated protein (MAP) kinases (Erk1 and Erk2, respectively) play a critical role in the regulation of cell growth and differentiation (12-15). MAP kinases are activated by a wide variety of extracellular signals including growth and neurotrophic factors, cytokines, hormones, and neurotransmitters. Activation of MAP kinases occur through phosphorylation of Thr202/Tyr204 on human Erk1 and Thr185/Tyr187 on human Erk2 at the sequence T*EY* by a pair of upstream MAP kinase kinases (MEK1/2) (16,17). Erk proteins are negatively regulated by a family of dual specificity (Thr/Tyr) MAPK phosphatases, known as DUSPs or MKPs (18), along with MEK inhibitors such as U0126 and PD98059. Erk dependent phosphorylation of TSC2 at Ser663 leads to the functional inactivation of the TSC1/TSC2 inhibitory complex, and subsequent downstream activation of S6 ribosomal protein through the mTORC1/p70 S6K signaling cascade (19).
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- Burgering, B.M. and Coffer, P.J. (1995) Nature 376, 599-602.
- Franke, T.F. et al. (1995) Cell 81, 727-36.
- Alessi, D.R. et al. (1996) EMBO J 15, 6541-51.
- Sarbassov, D.D. et al. (2005) Science 307, 1098-101.
- Myers, M.P. et al. (1998) Proc Natl Acad Sci U S A 95, 13513-8.
- Vlahos, C.J. et al. (1994) J Biol Chem 269, 5241-8.
- Peterson, R.T. and Schreiber, S.L. (1998) Curr Biol 8, R248-50.
- Jefferies, H.B. et al. (1997) EMBO J 16, 3693-704.
- Ferrari, S. et al. (1991) J Biol Chem 266, 22770-5.
- Flotow, H. and Thomas, G. (1992) J Biol Chem 267, 3074-8.
- Marshall, C.J. (1995) Cell 80, 179-85.
- Hunter, T. (1995) Cell 80, 225-36.
- Hill, C.S. and Treisman, R. (1995) Cell 80, 199-211.
- Cowley, S. et al. (1994) Cell 77, 841-52.
- Sturgill, T.W. et al. (1988) Nature 334, 715-8.
- Payne, D.M. et al. (1991) EMBO J 10, 885-92.
- Owens, D.M. and Keyse, S.M. (2007) Oncogene 26, 3203-13.
- Ma, L. et al. (2005) Cell 121, 179-93.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Applications Key
IF-IC: Immunofluorescence (Immunocytochemistry)
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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