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  • 名 称:
    磷酸化 p70 S6 Kinase (Thr389/412) 抗体
  • 货 号:
    AF3228
  • 来 源:
    Rabbit
  • 应 用:
    WB,IHC,IF/ICC,ELISA
  • 反 应:
    Human, Mouse, Rat, Pig
  • 预 测:
    Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(100%), Xenopus(100%)
  • 蛋白号:
  • 分子量:
    70kDa
  • 浓 度:
    1mg/ml
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产品信息

Alternative Names:Expand▼

70 kDa ribosomal protein S6 kinase 1; KS6B1_HUMAN; p70 alpha; P70 beta 1; p70 ribosomal S6 kinase alpha; p70 ribosomal S6 kinase beta 1; p70 S6 kinase alpha; P70 S6 Kinase; p70 S6 kinase, alpha 1; p70 S6 kinase, alpha 2; p70 S6K; p70 S6K-alpha; p70 S6KA; p70(S6K) alpha; p70(S6K)-alpha; p70-alpha; p70-S6K 1; p70-S6K; P70S6K; P70S6K1; p70S6Kb; PS6K; Ribosomal protein S6 kinase 70kDa polypeptide 1; Ribosomal protein S6 kinase beta 1; Ribosomal protein S6 kinase beta-1; Ribosomal protein S6 kinase I; RPS6KB1; S6K; S6K-beta-1; S6K1; Serine/threonine kinase 14 alpha; Serine/threonine-protein kinase 14A; STK14A;

Applications:

WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500, ELISA(peptide) 1:20000-1:40000

Reactivity:

Human, Mouse, Rat, Pig

Predicted Reactivity:

Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%), Chicken(100%), Xenopus(100%)

Source:

Rabbit

Clonality:

Polyclonal

Purification:

The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.

Specificity:

Phospho-p70 S6 Kinase (Thr389/412) Antibody detects endogenous levels of p70 S6 Kinase only when phosphorylated at Threonine 389/412.

Format:

Liquid

Concentration:

1mg/ml

Storage Condition and Buffer:

Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.Store at -20 °C.Stable for 12 months from date of receipt.

免疫原

Immunogen:

A synthesized peptide derived from human p70 S6 Kinase around the phosphorylation site of Thr389/412.

Uniprot:



>>Visit The Human Protein Atlas

Gene id:

Molecular Weight:

Observed Mol.Wt.: 70kDa.
Predicted Mol.Wt.: 60kDa.

Subcellular Location:

Cytoplasm; Nucleus. Cytoplasm and Cell junction > synapse > synaptosome. Mitochondrion outer membrane.

Tissue Specificity:

Widely expressed.

Description:

This gene encodes a member of the RSK (ribosomal S6 kinase) family of serine/threonine kinases. This kinase contains 2 non-identical kinase catalytic domains and phosphorylates several residues of the S6 ribosomal protein.

Sequence:
        10         20         30         40         50
MRRRRRRDGF YPAPDFRDRE AEDMAGVFDI DLDQPEDAGS EDELEEGGQL
60 70 80 90 100
NESMDHGGVG PYELGMEHCE KFEISETSVN RGPEKIRPEC FELLRVLGKG
110 120 130 140 150
GYGKVFQVRK VTGANTGKIF AMKVLKKAMI VRNAKDTAHT KAERNILEEV
160 170 180 190 200
KHPFIVDLIY AFQTGGKLYL ILEYLSGGEL FMQLEREGIF MEDTACFYLA
210 220 230 240 250
EISMALGHLH QKGIIYRDLK PENIMLNHQG HVKLTDFGLC KESIHDGTVT
260 270 280 290 300
HTFCGTIEYM APEILMRSGH NRAVDWWSLG ALMYDMLTGA PPFTGENRKK
310 320 330 340 350
TIDKILKCKL NLPPYLTQEA RDLLKKLLKR NAASRLGAGP GDAGEVQAHP
360 370 380 390 400
FFRHINWEEL LARKVEPPFK PLLQSEEDVS QFDSKFTRQT PVDSPDDSTL
410 420 430 440 450
SESANQVFLG FTYVAPSVLE SVKEKFSFEP KIRSPRRFIG SPRTPVSPVK
460 470 480 490 500
FSPGDFWGRG ASASTANPQT PVEYPMETSG IEQMDVTMSG EASAPLPIRQ
510 520
PNSGPYKKQA FPMISKRPEH LRMNL

研究背景

Function:

Serine/threonine-protein kinase that acts downstream of mTOR signaling in response to growth factors and nutrients to promote cell proliferation, cell growth and cell cycle progression. Regulates protein synthesis through phosphorylation of EIF4B, RPS6 and EEF2K, and contributes to cell survival by repressing the pro-apoptotic function of BAD. Under conditions of nutrient depletion, the inactive form associates with the EIF3 translation initiation complex. Upon mitogenic stimulation, phosphorylation by the mammalian target of rapamycin complex 1 (mTORC1) leads to dissociation from the EIF3 complex and activation. The active form then phosphorylates and activates several substrates in the pre-initiation complex, including the EIF2B complex and the cap-binding complex component EIF4B. Also controls translation initiation by phosphorylating a negative regulator of EIF4A, PDCD4, targeting it for ubiquitination and subsequent proteolysis. Promotes initiation of the pioneer round of protein synthesis by phosphorylating POLDIP3/SKAR. In response to IGF1, activates translation elongation by phosphorylating EEF2 kinase (EEF2K), which leads to its inhibition and thus activation of EEF2. Also plays a role in feedback regulation of mTORC2 by mTORC1 by phosphorylating RICTOR, resulting in the inhibition of mTORC2 and AKT1 signaling. Mediates cell survival by phosphorylating the pro-apoptotic protein BAD and suppressing its pro-apoptotic function. Phosphorylates mitochondrial URI1 leading to dissociation of a URI1-PPP1CC complex. The free mitochondrial PPP1CC can then dephosphorylate RPS6KB1 at Thr-412, which is proposed to be a negative feedback mechanism for the RPS6KB1 anti-apoptotic function. Mediates TNF-alpha-induced insulin resistance by phosphorylating IRS1 at multiple serine residues, resulting in accelerated degradation of IRS1. In cells lacking functional TSC1-2 complex, constitutively phosphorylates and inhibits GSK3B. May be involved in cytoskeletal rearrangement through binding to neurabin. Phosphorylates and activates the pyrimidine biosynthesis enzyme CAD, downstream of MTOR (PubMed:11500364, PubMed:12801526, PubMed:14673156, PubMed:15071500, PubMed:15341740, PubMed:16286006, PubMed:17052453, PubMed:17053147, PubMed:17936702, PubMed:18952604, PubMed:19085255, PubMed:19720745, PubMed:19935711, PubMed:19995915, PubMed:23429703). Following activation by mTORC1, phosphorylates EPRS and thereby plays a key role in fatty acid uptake by adipocytes and also most probably in interferon-gamma-induced translation inhibition (PubMed:28178239).

Post-translational Modifications:

Phosphorylation at Thr-412 is regulated by mTORC1. The phosphorylation at this site is maintained by an agonist-dependent autophosphorylation mechanism (By similarity). Activated by phosphorylation at Thr-252 by PDPK1. Dephosphorylation by PPP1CC at Thr-412 in mitochondrion.

Subcellular Location:

Mitochondrion;Nucleus;

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionGraphics by Christian Stolte

Subunit Structure:

Interacts with PPP1R9A/neurabin-1 (By similarity). Interacts with RPTOR (PubMed:12150926). Interacts with IRS1 (PubMed:18952604). Interacts with EIF3B and EIF3C (PubMed:16286006). Interacts with TRAF4 (PubMed:12801526). Interacts with POLDIP3 (PubMed:15341740). Interacts (via N-terminus) with IER5 (PubMed:26496226).

Similarity:

The autoinhibitory domain is believed to block phosphorylation within the AGC-kinase C-terminal domain and the activation loop.The TOS (TOR signaling) motif is essential for activation by mTORC1.Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. S6 kinase subfamily.

研究领域

Research Fields:

· Cellular Processes > Transport and catabolism > Autophagy - animal.(View pathway)
· Environmental Information Processing > Signal transduction > ErbB signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > mTOR signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > Apelin signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > TGF-beta signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > HIF-1 signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > AMPK signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway.(View pathway)
· Human Diseases > Cancers: Specific types > Pancreatic cancer.(View pathway)
· Human Diseases > Cancers: Overview > Pathways in cancer.(View pathway)
· Human Diseases > Cancers: Specific types > Gastric cancer.(View pathway)
· Human Diseases > Cancers: Specific types > Colorectal cancer.(View pathway)
· Human Diseases > Cancers: Overview > Proteoglycans in cancer.
· Human Diseases > Cancers: Specific types > Hepatocellular carcinoma.(View pathway)
· Human Diseases > Cancers: Specific types > Acute myeloid leukemia.(View pathway)
· Human Diseases > Cancers: Specific types > Breast cancer.(View pathway)
· Human Diseases > Endocrine and metabolic diseases > Insulin resistance.
· Human Diseases > Cancers: Overview > Choline metabolism in cancer.(View pathway)
· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.
· Human Diseases > Drug resistance: Antineoplastic > EGFR tyrosine kinase inhibitor resistance.
· Human Diseases > Drug resistance: Antineoplastic > Endocrine resistance.
· Organismal Systems > Immune system > Fc gamma R-mediated phagocytosis.(View pathway)
· Organismal Systems > Endocrine system > Insulin signaling pathway.(View pathway)
· Organismal Systems > Aging > Longevity regulating pathway.(View pathway)
· Organismal Systems > Aging > Longevity regulating pathway - multiple species.(View pathway)

Western blot analysis of p70 S6 Kinase phosphorylation expression in Insulin treated Hela whole cell lysates,The lane on the left was treated with the antigen-specific peptide.
Western blot analysis of Phospho-p70 S6 Kinase (Thr389/412) using various lysates Lanes 1 - 2: Merged signal (red and green). Green - AF3228 observed at 70 kDa. Red - loading control, T0023, observed at 55 kDa. Blots were developed with Goat Anti-Rabbit IgG(H+L) FITC–conjugated (S0008) and Goat Anti-Mouse IgG(H+L) Alexa Fluor 594–conjugated (S0005) secondary antibodies
AF3228 at 1/200 staining human liver cancer tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22°C. An HRP conjugated goat anti-rabbit antibody was used as the secondary.
AF3228 staining 293 cells by ICC/IF. Cells were fixed with PFA and permeabilized in 0.1% saponin prior to blocking in 10% serum for 45 minutes at 37°C. The primary antibody was diluted 1/400 and incubated with the sample for 1 hour at 37°C. A Alexa Fluor 594 conjugated goat polyclonal to rabbit IgG (H+L), diluted 1/600 was used as secondary antibody.
ELISA analysis of AF3228 showing specificity to Phospho-p70 S6 Kinase (Thr389/412) peptide. Peptides concentration: 1ug/ml.
P-peptide: phospho-peptide; N-peptide: non-phospho-peptide.

Reference Citations:

1). Shen F et al. CEMIP promotes ovarian cancer development and progression via the PI3K/AKT signaling pathway. Biomed Pharmacother 2019 Mar 26;114:108787 (PubMed: 30925458) [IF=3.743]

2). Lin H et al. Rapamycin Supplementation May Ameliorate Erectile Function in Rats With Streptozotocin-Induced Type 1 Diabetes by Inducing Autophagy and Inhibiting Apoptosis, Endothelial Dysfunction, and Corporal Fibrosis. J Sex Med 2018 Sep;15(9):1246-1259 (PubMed: 30224017) [IF=3.649]

3). Wu Z et al. Traditional Chinese Medicine CFF-1 induced cell growth inhibition, autophagy, and apoptosis via inhibiting EGFR-related pathways in prostate cancer. Cancer Med 2018 Apr;7(4):1546-1559 (PubMed: 29533017) [IF=3.357]

4). Zheng L et al. Supplementation of branched-chain amino acids to a reduced-protein diet improves growth performance in piglets: involvement of increased feed intake and direct muscle growth-promoting effect. Br J Nutr 2016 Jun;115(12):2236-45 (PubMed: 27079773) [IF=3.319]

Application: WB    Species:pig;    Sample:Not available

Fig. 4. Abundance of phosphorylated and total (A) ribosomal protein S6 kinase 1 (S6K1) and (B) mammalian target of rapamycin (mTOR) proteins in the hypothalamus of piglets fed positive control (PC, ) diet, reduced-protein negative control (NC, ) diet or two NC diets supplemented with a 1- or 2-fold dose of BCAA (test 1 ( ) and test 2 ( ), respectively) in Expt 1. Western blotting method was used. Values were normalised using β-actin or relative total protein as an internal control. Data are expressed relative to expressions in the PC group; values are means (n 4–6/group), with their standard errors represented by vertical bars. a,b Mean values with unlike letters were significantly different (P <0·05).


5). Xu W et al. TAZ inhibition restores sensitivity of cisplatin via AKT/mTOR signaling in lung adenocarcinoma. Oncol Rep 2017 Sep;38(3):1815-1821 (PubMed: 28737828)

6). Sun X et al. Hepatic conditional knockout of ATF6 exacerbates liver metabolic damage by repressing autophage through MTOR pathway. Biochem Biophys Res Commun 2018 Oct 20;505(1):45-50 (PubMed: 30236984)

7). Wang D et al. The effects of reduced dietary protein level on amino acid transporters and mTOR signaling pathway in pigs. Biochem Biophys Res Commun 2017 Apr 1;485(2):319-327 (PubMed: 28235480)

8). Wu Z et al. EGFR‑associated pathways involved in traditional Chinese medicine (TCM)‑1‑induced cell growth inhibition, autophagy and apoptosis in prostate cancer. Mol Med Rep 2018 Jun;17(6):7875-7885 (PubMed: 29620175)

9). Wu Z et al. EGFR‑associated pathways involved in traditional Chinese medicine (TCM)‑1‑induced cell growth inhibition, autophagy and apoptosis in prostate cancer. Mol Med Rep 2018 Jun;17(6):7875-7885 (PubMed: 29620175)

10). Wu D et al. Olaquindox disrupts tight junction integrity and cytoskeleton architecture in mouse Sertoli cells. Oncotarget 2017 Aug 16;8(51):88630-88644 (PubMed: 29179463)

11). Jin J et al. Rapamycin Reduces Podocyte Apoptosis and is Involved in Autophagy and mTOR/ P70S6K/4EBP1 Signaling. Cell Physiol Biochem 2018;48(2):765-772 (PubMed: 30025409)

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Catalog Number :

AF3228-BP
(Blocking peptide available as AF3228-BP)

Price/Size :

¥2000/1mg.
Tips: For phospho antibody, we provide phospho peptide(0.5mg) and non-phospho peptide(0.5mg).

Function :

Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. This mechanism is useful when non-specific binding is an issue, for example, in Western blotting (immunoblot) and immunohistochemistry (IHC). By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific; Specific binding will be absent from the western blot or immunostaining performed with the neutralized antibody.

Format and storage :

Synthetic peptide was lyophilized with 100% acetonitrile and is supplied as a powder. Reconstitute with 0.1 ml DI water for a final concentration of 10 mg/ml.The purity is >90%,tested by HPLC and MS.Storage Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C.

Precautions :

This product is for research use only. Not for use in diagnostic or therapeutic procedures.

Xenopus
100%
Chicken
100%
Rabbit
100%
Pig
100%
Dog
100%
Bovine
100%
Sheep
100%
Horse
100%
Zebrafish
0%
High similarity Medium similarity Low similarity No similarity
P23443 as Substrate
Site PTM Type Enzyme
S40 Phosphorylation
S53 Phosphorylation Q9HC98 (NEK6)
K85 Ubiquitination
K99 Ubiquitination
K104 Ubiquitination
K118 Ubiquitination
S243 Phosphorylation
T248 Phosphorylation
T250 Phosphorylation
T252 Phosphorylation P42336 (PIK3CA) , O15530 (PDPK1)
T256 Phosphorylation
S278 Phosphorylation
K304 Acetylation
K364 Ubiquitination
K370 Ubiquitination
S371 Phosphorylation
T389 Phosphorylation
T390 Phosphorylation P42345 (MTOR)
S394 Phosphorylation P06493 (CDK1) , P42345 (MTOR)
T399 Phosphorylation
S403 Phosphorylation Q9HC98 (NEK6)
T412 Phosphorylation O94768 (STK17B) , O15530 (PDPK1) , Q8TDX7 (NEK7) , Q9HC98 (NEK6) , P42336 (PIK3CA) , P42345 (MTOR) , P23443 (RPS6KB1)
Y413 Phosphorylation
S417 Phosphorylation
S421 Phosphorylation
K425 Ubiquitination
S427 Phosphorylation
S434 Phosphorylation P06493 (CDK1) , P42345 (MTOR) , P45984 (MAPK9) , P27361 (MAPK3) , P45983 (MAPK8) , P28482 (MAPK1)
S441 Phosphorylation
T444 Phosphorylation P06493 (CDK1) , P28482 (MAPK1)
S447 Phosphorylation P28482 (MAPK1) , P06493 (CDK1) , P42345 (MTOR)
K450 Ubiquitination
S452 Phosphorylation
S462 Phosphorylation
T470 Phosphorylation
K516 Acetylation
R522 Methylation
P23443 as PTM Enzyme
Substrate Site Source
O00418 (EEF2K) S366 Uniprot
O60825 (PFKFB2) S466 Uniprot
O94763 (URI1) S372 Uniprot
P03372 (ESR1) S167 Uniprot
P04637 (TP53) S392 Uniprot
P04792 (HSPB1) S15 Uniprot
P04792 (HSPB1) S78 Uniprot
P04792 (HSPB1) S82 Uniprot
P08151 (GLI1) S84 Uniprot
P10636-8 (MAPT) T212 Uniprot
P10636-8 (MAPT) S214 Uniprot
P10636-8 (MAPT) S262 Uniprot
P23443 (RPS6KB1) T412 Uniprot
P23588-1 (EIF4B) S422 Uniprot
P27708 (CAD) S1859 Uniprot
P35568 (IRS1) S270 Uniprot
P35568 (IRS1) S307 Uniprot
P35568 (IRS1) S527 Uniprot
P35568 (IRS1) S636 Uniprot
P35568 (IRS1) S1101 Uniprot
P42345 (MTOR) T2446 Uniprot
P42345 (MTOR) S2448 Uniprot
P49841 (GSK3B) S9 Uniprot
P62753 (RPS6) S235 Uniprot
P62753 (RPS6) S236 Uniprot
P62753 (RPS6) S240 Uniprot
P62753 (RPS6) S244 Uniprot
P62753 (RPS6) S247 Uniprot
P78371 (CCT2) S260 Uniprot
Q05195 (MXD1) S145 Uniprot
Q06787 (FMR1) S500 Uniprot
Q09161 (NCBP1) S7 Uniprot
Q09161 (NCBP1) T21 Uniprot
Q09161 (NCBP1) S22 Uniprot
Q15831-1 (STK11) S428 Uniprot
Q16873 (LTC4S) S36 Uniprot
Q53EL6 (PDCD4) S67 Uniprot
Q6R327 (RICTOR) T1135 Uniprot
Q8TB45 (DEPTOR) S286 Uniprot
Q8TB45 (DEPTOR) S287 Uniprot
Q8TB45 (DEPTOR) S291 Uniprot
Q92519 (TRIB2) S83 Uniprot
Q92934 (BAD) S99 Uniprot
Q9BY77 (POLDIP3) S383 Uniprot
Q9BY77 (POLDIP3) S385 Uniprot
Q9BYV9 (BACH2) S521 Uniprot
Q9H3D4 (TP63) S477 Uniprot
Q9H3D4 (TP63) T491 Uniprot
Q9H3D4 (TP63) S560 Uniprot
Q9UN36 (NDRG2) S332 Uniprot
Q9UN36 (NDRG2) S350 Uniprot
IMPORTANT: For western blots, incubate membrane with diluted antibody in 5% w/v milk , 1X TBS, 0.1% Tween®20 at 4°C with gentle shaking, overnight.

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