mTOR (Ser2481) Polyclonal Antibody

$Formalin-fixed and paraffin embedded rat brain labeled with Rabbit Anti-mTOR (Ser2481) Polyclonal Antibody, Unconjugated(bs-3495R) 1:200 followed by conjugation to the secondary antibody and DAB staining\\n$

Formalin-fixed and paraffin embedded rat brain labeled with Rabbit Anti-mTOR (Ser2481) Polyclonal Antibody, Unconjugated(bs-3495R) 1:200 followed by conjugation to the secondary antibody and DAB staining\\n

Mouse splenocytes probed with Rabbit Anti-mTOR (Ser2481) Polyclonal Antibody, Unconjugated (bs-3495R) (green) at 1:50 for 30 minutes followed by a x conjugated secondary antibody compared to unstained cells (blue), secondary only(light blue), and isotype control(orange).

HepG2 lysates probed with mTOR (Ser2481) Polyclonal Antibody, Unconjugated (bs-3495R) at 1:300 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:10000 for 60 min at 37˚C.

$Paraformaldehyde-fixed, paraffin embedded Mouse brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with mTOR (Ser2481) Polyclonal Antibody, Unconjugated (bs-3495R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Mouse brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with mTOR (Ser2481) Polyclonal Antibody, Unconjugated (bs-3495R) at 1:200 overnight at 4\u00b0C, DAB staining.

$A431 cells were\u00a0fixed with 4% PFA for 10min at room temperature,permeabilized with\u00a090% ice-cold methanol for 20 min at -20\u2103,\u00a0and incubated in 5% BSA blocking buffer\u00a0for 30 min at room temperature. Cells were then stained with Phospho-mTOR (Ser2481) Polyclonal Antibody(bs-3495R)at 1:100 dilution\u00a0in blocking buffer and\u00a0incubated for 30 min at\u00a0room temperature,\u00a0washed twice with 2%BSA in PBS,\u00a0followed by\u00a0secondary antibody incubation\u00a0for 40 min\u00a0at\u00a0room temperature. Acquisitions of 20,000 events were performed.\u00a0Cells stained with primary antibody\u00a0(green), and isotype control (orange).$

A431 cells were\u00a0fixed with 4% PFA for 10min at room temperature,permeabilized with\u00a090% ice-cold methanol for 20 min at -20\u2103,\u00a0and incubated in 5% BSA blocking buffer\u00a0for 30 min at room temperature. Cells were then stained with Phospho-mTOR (Ser2481) Polyclonal Antibody(bs-3495R)at 1:100 dilution\u00a0in blocking buffer and\u00a0incubated for 30 min at\u00a0room temperature,\u00a0washed twice with 2%BSA in PBS,\u00a0followed by\u00a0secondary antibody incubation\u00a0for 40 min\u00a0at\u00a0room temperature. Acquisitions of 20,000 events were performed.\u00a0Cells stained with primary antibody\u00a0(green), and isotype control (orange).

$A431 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37\u00b0C for 20 min; Antibody incubation with (Phospho-mTOR (Ser2481)) polyclonal Antibody, Unconjugated (bs-3495R) 1:100, 90 minutes at 37\u00b0C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37\u00b0C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei$

A431 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37\u00b0C for 20 min; Antibody incubation with (Phospho-mTOR (Ser2481)) polyclonal Antibody, Unconjugated (bs-3495R) 1:100, 90 minutes at 37\u00b0C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37\u00b0C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei

Applications

WB
ELISA
FCM
IHC-P
IHC-F
IF(IHC-P)
IF(IHC-F)
IF(ICC)

Reactivity

Human
Mouse
Rat

Predicted Reactivity

Dog
Sheep
Pig
Horse
Rabbit

Overview
Catalog #	bs-3495R
Product Name	mTOR (Ser2481) Polyclonal Antibody
Applications	WB, ELISA, FCM, IHC-P, IHC-F, IF(IHC-P), IF(IHC-F), IF(ICC)
Specificity	This phosphorylation site is homologous in the listed cross reactive species at the specified location.
Reactivity	Human, Mouse, Rat
Predicted Reactivity	Dog, Sheep, Pig, Horse, Rabbit
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	KLH conjugated synthetic phosphopeptide derived from human mTOR around the phosphorylation site of Ser2481
Modification Site	Ser2481
Clonality	Polyclonal
Isotype	IgG
Concentration	1ug/ul
Purification	Purified by Protein A.
Storage Buffer	0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage Condition	Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID	2475
Swiss Prot	P42345
Subcellular location	Cytoplasm, Cell membrane
Synonyms	FRAP; FRAP1; FRAP2; RAFT1; RAPT1; Serine/threonine-protein kinase mTOR; FK506-binding protein 12-rapamycin complex-associated protein 1; FKBP12-rapamycin complex-associated protein; Mammalian target of rapamycin; mTOR; Mechanistic target of rapamycin; Rapamycin and FKBP12 target 1; Rapamycin target protein 1
Background	Serine/threonine protein kinase which is a central regulator of cellular metabolism, growth and survival in response to hormones, growth factors, nutrients, energy and stress signals. MTOR directly or indirectly regulates the phosphorylation of at least 800 proteins. Functions as part of 2 structurally and functionally distinct signaling complexes mTORC1 and mTORC2 (mTOR complex 1 and 2). Activated mTORC1 up-regulates protein synthesis by phosphorylating key regulators of mRNA translation and ribosome synthesis. This includes phosphorylation of EIF4EBP1 and release of its inhibition toward the elongation initiation factor 4E (eiF4E). Moreover, phosphorylates and activates RPS6KB1 and RPS6KB2 that promote protein synthesis by modulating the activity of their downstream targets including ribosomal protein S6, eukaryotic translation initiation factor EIF4B, and the inhibitor of translation initiation PDCD4. Stimulates the pyrimidine biosynthesis pathway, both by acute regulation through RPS6KB1-mediated phosphorylation of the biosynthetic enzyme CAD, and delayed regulation, through transcriptional enhancement of the pentose phosphate pathway which produces 5-phosphoribosyl-1-pyrophosphate (PRPP), an allosteric activator of CAD at a later step in synthesis, this function is dependent on the mTORC1 complex. Regulates ribosome synthesis by activating RNA polymerase III-dependent transcription through phosphorylation and inhibition of MAF1 an RNA polymerase III-repressor. In parallel to protein synthesis, also regulates lipid synthesis through SREBF1/SREBP1 and LPIN1. To maintain energy homeostasis mTORC1 may also regulate mitochondrial biogenesis through regulation of PPARGC1A. mTORC1 also negatively regulates autophagy through phosphorylation of ULK1. Under nutrient sufficiency, phosphorylates ULK1 at 'Ser-758', disrupting the interaction with AMPK and preventing activation of ULK1. Also prevents autophagy through phosphorylation of the autophagy inhibitor DAP.
Application Dilution
WB	1:300-5000
ELISA	1:500-1000
FCM	1:20-100
IHC-P	1:200-400
IHC-F	1:100-500
IF(IHC-P)	1:50-200
IF(IHC-F)	1:50-200
IF(ICC)	1:50-200