Phospho-RPS6KB1 (T421+S424) Antibody

Datasheet
Code CSB-RA020470A421phHU
Size US$350
Uniprot No. P23443
Image
  • Western Blot
    Positive WB detected in:293 whole cell lysate(treated with Calyculin A or not)
    All lanes:Phospho-RPS6KB1 antibody at 0.93μg/ml
    Secondary
    Goat polyclonal to rabbit IgG at 1/50000 dilution
    Predicted band size: 70 KDa
    Observed band size: 70 KDa

  • Immunofluorescence staining of Hela cells with CSB-RA020470A421phHU at 1:100,counter-stained with DAPI. The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Rabbit IgG (H+L).

  • Immunoprecipitating Phospho-RPS6KB1 in Hela whole cell lysate
    Lane 1: Rabbit control IgG(1μg)instead of CSB-RA020470A421phHU in Hela whole cell lysate. For western blotting,a HRP-conjugated Protein G antibody was used as the secondary antibody (1/2000)
    Lane 2: CSB-RA020470A421phHU(3μg)+ Hela whole cell lysate(1mg)
    Lane 3: Hela whole cell lysate (20μg)

Protocols ELISA Protocol
Western Blotting(WB) Protocol
Immunofluorescence (IF) Protocol
Immunoprecipitation (IP) Protocol
Immunogen A synthesized peptide derived from Human Phospho-RPS6KB1 (T421+S424)
Species Reactivity Human
Tested Applications ELISA, WB, IF, IP; Recommended dilution: WB:1:500-1:5000, IF:1:20-1:200, IP:1:200-1:1000
Relevance 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.
Form Liquid
Conjugate Non-conjugated
Storage Buffer Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.
Purification Method Affinity-chromatography
Isotype Rabbit IgG
Clonality Monoclonal
Alias Ribosomal protein S6 kinase beta-1, 70 kDa ribosomal protein S6 kinase 1, P70S6K1, p70-S6K 1, Ribosomal protein S6 kinase I, Serine/threonine-protein kinase 14A, p70 ribosomal S6 kinase alpha, p70 S6 kinase alpha, p70 S6K-alpha, p70 S6KA, RPS6KB1, STK14A
Immunogen Species Homo sapiens (Human)
Research Area Cell Biology
Gene Names RPS6KB1
Storage Upon receipt, store at -20°C or -80°C. Avoid repeated freeze.
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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.
Subcellular Location Cell junction, synapse, synaptosome, Mitochondrion outer membrane, Mitochondrion
Protein Families Protein kinase superfamily, AGC Ser/Thr protein kinase family, S6 kinase subfamily
Tissue Specificity Widely expressed.
Database Links

HGNC: 10436

OMIM: 608938

KEGG: hsa:6198

STRING: 9606.ENSP00000225577

UniGene: Hs.463642

Pathway ErbB signaling pathway
HIF-1 signaling pathway
mTOR signaling pathway
PI3K-Akt signaling pathway
TGF-beta signaling pathway
Autophagy
Fc gamma R-mediated phagocytosis
AMPK signaling
Apelin signaling pathway
Insulin signaling pathway

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