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Rabbit Monoclonal KS6B1 phospho T389 antibody. Suitable for Dot, WB and reacts with Synthetic peptide - Human, Human samples.

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Images

Western blot - Anti-S6K1 (phospho T389) antibody [EPR24766-9] (AB323272), expandable thumbnail
  • Western blot - Anti-S6K1 (phospho T389) antibody [EPR24766-9] (AB323272), expandable thumbnail
  • Dot Blot - Anti-S6K1 (phospho T389) antibody [EPR24766-9] (AB323272), expandable thumbnail

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
DotWBICC/IFIHC-P
Human
Expected
Tested
Not recommended
Not recommended
Mouse
Not recommended
Not recommended
Not recommended
Not recommended
Rat
Not recommended
Not recommended
Not recommended
Not recommended
Synthetic peptide - Human
Tested
Not recommended
Not recommended
Not recommended

Tested
Tested

Species
Synthetic peptide - Human
Dilution info
1/1000
Notes

-

Expected
Expected

Species
Human
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Mouse, Rat
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/1000
Notes

-

Not recommended
Not recommended

Species
Mouse, Rat, Synthetic peptide - Human
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Human, Mouse, Rat, Synthetic peptide - Human
Dilution info
-
Notes

-

Not recommended
Not recommended

Species
Human, Mouse, Rat, Synthetic peptide - Human
Dilution info
-
Notes

-

Target data

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 (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:22017876, PubMed:23429703, PubMed:28178239). Regulates protein synthesis through phosphorylation of EIF4B, RPS6 and EEF2K, and contributes to cell survival by repressing the pro-apoptotic function of BAD (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:22017876, PubMed:23429703, PubMed:28178239). Under conditions of nutrient depletion, the inactive form associates with the EIF3 translation initiation complex (PubMed:16286006). Upon mitogenic stimulation, phosphorylation by the mechanistic target of rapamycin complex 1 (mTORC1) leads to dissociation from the EIF3 complex and activation (PubMed:16286006). 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 (PubMed:16286006). Also controls translation initiation by phosphorylating a negative regulator of EIF4A, PDCD4, targeting it for ubiquitination and subsequent proteolysis (PubMed:17053147). Promotes initiation of the pioneer round of protein synthesis by phosphorylating POLDIP3/SKAR (PubMed:15341740). In response to IGF1, activates translation elongation by phosphorylating EEF2 kinase (EEF2K), which leads to its inhibition and thus activation of EEF2 (PubMed:11500364). Also plays a role in feedback regulation of mTORC2 by mTORC1 by phosphorylating RICTOR, resulting in the inhibition of mTORC2 and AKT1 signaling (PubMed:19720745, PubMed:19935711, PubMed:19995915). Also involved in feedback regulation of mTORC1 and mTORC2 by phosphorylating DEPTOR (PubMed:22017876). Mediates cell survival by phosphorylating the pro-apoptotic protein BAD and suppressing its pro-apoptotic function (By similarity). Phosphorylates mitochondrial URI1 leading to dissociation of a URI1-PPP1CC complex (PubMed:17936702). 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 (PubMed:17936702). Mediates TNF-alpha-induced insulin resistance by phosphorylating IRS1 at multiple serine residues, resulting in accelerated degradation of IRS1 (PubMed:18952604). In cells lacking functional TSC1-2 complex, constitutively phosphorylates and inhibits GSK3B (PubMed:17052453). May be involved in cytoskeletal rearrangement through binding to neurabin (By similarity). Phosphorylates and activates the pyrimidine biosynthesis enzyme CAD, downstream of MTOR (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).

Alternative names

STK14A, RPS6KB1, Ribosomal protein S6 kinase beta-1, S6K-beta-1, S6K1, 70 kDa ribosomal protein S6 kinase 1, Ribosomal protein S6 kinase I, Serine/threonine-protein kinase 14A, p70 ribosomal S6 kinase alpha, P70S6K1, p70-S6K 1, p70 S6 kinase alpha, p70 S6K-alpha, p70 S6KA

Rabbit Monoclonal KS6B1 phospho T389 antibody. Suitable for Dot, WB and reacts with Synthetic peptide - Human, Human samples.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
EPR24766-9
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

3 product images

  • Western blot - Anti-S6K1 (phospho T389) antibody [EPR24766-9] (ab323272), expandable thumbnail

    Western blot - Anti-S6K1 (phospho T389) antibody [EPR24766-9] (ab323272)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    The identity of the higher MW band at approximately 150 kDa (in lane 2) is unknown.

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

    In Western blot, Anti-RPS6KB1 antibody [E343] (Anti-S6K1 antibody [E343] ab32529) staining at 1/1000 dilution.

    All lanes: Western blot - Anti-S6K1 (phospho T389) antibody [EPR24766-9] (ab323272) at 1/1000 dilution

    Lane 1: Untreated HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 50 µg

    Lane 2: Hela treated with 100nM Calycin A for 30min whole cell lysate at 50 µg

    Lane 3: Hela treated with 100nM Calycin A for 30min whole cell lysate (alkaline phosphatase treated membrane) at 50 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Observed band size: 60-80 kDa, 36 kDa

    Exposure time: 37s

  • Western blot - Anti-S6K1 (phospho T389) antibody [EPR24766-9] (ab323272), expandable thumbnail

    Western blot - Anti-S6K1 (phospho T389) antibody [EPR24766-9] (ab323272)

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    Performed under reducing conditions.

    In Western blot, ab323272 was shown to bind specifically to RPS6KB1. Target of interest was observed at 60–80kDa in wild-type HAP1 cell lysates (lane 2) with no signal observed at this size in RPS6KB1 knockout cell line (lane 4).

    The identity of the higher MW band at approximately 150 kDa (in lanes 2/4) is unknown.

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

    In Western blot, Anti-RPS6KB1 antibody [E343] (Anti-S6K1 antibody [E343] ab32529) staining at 1/1000 dilution.

    All lanes: Western blot - Anti-S6K1 (phospho T389) antibody [EPR24766-9] (ab323272) at 1/1000 dilution

    Lane 1: HAP1(HOWT01) (human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 50 µg

    Lane 2: Parental HAP1(HOWT01) treated with 100nM Calycin A for 30min whole cell lysate at 50 µg

    Lane 3: Untreated RPS6KB1 knockout HAP1 whole cell lysate at 50 µg

    Lane 4: RPS6KB1 knockout HAP1 treated with 100nM Calycin A whole cell lysate at 50 µg

    Lane 5: Untreated parental HAP1(HOWT01) (human chronic myelogenous leukemia near-haploid cell) whole cell lysate (alkaline phosphatase treated membrane) at 50 µg

    Lane 6: Parental HAP1(HOWT01) treated with 100nM Calyculin A for 30min whole cell lysate (alkaline phosphatase treated membrane) at 50 µg

    Lane 7: Untreated RPS6KB1 knockout HAP1 whole cell lysate (alkaline phosphatase treated membrane) at 50 µg

    Lane 8: RPS6KB1 knockout HAP1 treated with 100nM Calyculin A whole cell lysate (alkaline phosphatase treated membrane) at 50 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

    Performed under reducing conditions.

    Observed band size: 60-80 kDa, 36 kDa

    Exposure time: 180s

  • Dot Blot - Anti-S6K1 (phospho T389) antibody [EPR24766-9] (ab323272), expandable thumbnail

    Dot Blot - Anti-S6K1 (phospho T389) antibody [EPR24766-9] (ab323272)

    Dot blot analysis of S6K1 (phospho T389) using ab323272 at 1:1000 (0.48 ug/ml) followed by a VeriBlot for IP secondary antibody (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1:5000 dilution.

    Lane1: Ribosomal protein S6 kinase beta-1/p70 S6K (phospho T389) peptide

    Lane2: Ribosomal protein S6 kinase beta-1/p70 S6K non-phospho peptide

    Lane3: Ribosomal protein S6 kinase beta-1/p70 S6K (phospho S393) peptide

    Lane4: Ribosomal protein S6 kinase beta-1/p70 S6K (phospho Y390) peptide

    Exposure time: 180 seconds.

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    All lanes: Dot Blot - Anti-S6K1 (phospho T389) antibody [EPR24766-9] (ab323272) at 1/1000 dilution

    Lane 1: Ribosomal protein S6 kinase beta-1/p70 S6K (phospho T389) peptide

    Lane 2: Ribosomal protein S6 kinase beta-1/p70 S6K non-phospho peptide

    Lane 3: Ribosomal protein S6 kinase beta-1/p70 S6K (phospho S393) peptide

    Lane 4: Ribosomal protein S6 kinase beta-1/p70 S6K (phospho Y390) peptide

    Secondary

    All lanes: Dot Blot - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

    Exposure time: 180s

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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