Phospho-Sin1 (Thr86) (D4U9L) Rabbit mAb (#14716) Datasheet Without Images Cell Signaling Technology

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:

WB, IP

REACTIVITY:

H M

SENSITIVITY:

Endogenous

MW (kDa):

78, 74

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q9BPZ7

Entrez-Gene Id:

79109

Product Information

Product Usage Information

Application	Dilution
Western Blotting	1:1000
Immunoprecipitation	1:50

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.

Specificity / Sensitivity

Phospho-Sin1 (Thr86) (D4U9L) Rabbit mAb recognizes endogenous levels of Sin1 protein only when phosphorylated at Thr86.

Species Reactivity:

Human, Mouse

Species predicted to react based on 100% sequence homology

Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding phosphorylated Thr86 of human Sin1 protein.

Background

Cell growth is a fundamental biological process whereby cells accumulate mass and increase in size. The mammalian TOR (mTOR) pathway regulates growth by coordinating energy and nutrient signals with growth factor-derived signals (1). mTOR is a large protein kinase that is a component of two different complexes. The mTOR complex 1 (mTORC1), a target of rapamycin, contains mTOR, GβL, and raptor. mTORC2, insensitive to rapamycin, includes mTOR, GβL, Sin1, and rictor (1). The mTORC2 complex phosphorylates Ser473 of Akt/PKB in vitro (2). This phosphorylation is essential for full Akt/PKB activation. Furthermore, rictor siRNA knockdown inhibits Ser473 phosphorylation in 3T3-L1 adipocytes (3). mTORC2 has also been shown to phosphorylate the rapamycin-resistant mutants of S6K1, another effector of mTOR (4). In addition, phosphorylation of Sin1 at Thr86 by Akt/PKB was shown to regulate the activity of mTORC2 in adipocytes upon stimulation by growth factors (5).

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation

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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