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Rabbit Multiclonal KS6B1 phospho T421 + S424 antibody. Suitable for WB, ICC/IF and reacts with Human samples. Immunogen corresponding to Synthetic Peptide within Human RPS6KB1 phospho T421 + S424.

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Images

Immunocytochemistry/ Immunofluorescence - Anti-p70 S6 Kinase (phospho T421 + S424) antibody [RP23040018] (AB308099), expandable thumbnail
  • Western blot - Anti-p70 S6 Kinase (phospho T421 + S424) antibody [RP23040018] (AB308099), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-p70 S6 Kinase (phospho T421 + S424) antibody [RP23040018] (AB308099), expandable thumbnail
  • Western blot - Anti-p70 S6 Kinase (phospho T421 + S424) antibody [RP23040018] (AB308099), expandable thumbnail

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

Preservative: 0.09% Sodium azide
Constituents: 99.91% PBS

Form
Liquid
Clonality
Multiclonal

Immunogen

  • Synthetic Peptide within Human RPS6KB1 phospho T421 + S424. Database link P23443

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBICC/IF
Human
Tested
Tested

Tested
Tested

Species
Human
Dilution info
0.5-1 µg/mL
Notes

-

Tested
Tested

Species
Human
Dilution info
1/200
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 Multiclonal KS6B1 phospho T421 + S424 antibody. Suitable for WB, ICC/IF and reacts with Human samples. Immunogen corresponding to Synthetic Peptide within Human RPS6KB1 phospho T421 + S424.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Multiclonal
Immunogen
  • Synthetic Peptide within Human RPS6KB1 phospho T421 + S424. Database link P23443
Clone number
RP23040018
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

What are recombinant multiclonals?
Recombinant multiclonals are a mixture of recombinant antibodies co-expressed from a library of heavy and light chains. They offer several advantages including:

  • - The sensitivity of polyclonal antibodies by recognising multiple epitopes
  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

View our range of recombinant multiclonal 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.

4 product images

  • Immunocytochemistry/ Immunofluorescence - Anti-p70 S6 Kinase (phospho T421 + S424) antibody [RP23040018] (ab308099), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-p70 S6 Kinase (phospho T421 + S424) antibody [RP23040018] (ab308099)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed serum-starved HeLa cells treated with insulin (100 ng/mL, 15 min) labeling p70 S6 Kinase (phospho T421 + S424) with ab308099, followed by Alexa Fluor® 488 Goat anti-Rabbit IgG secondary antibody (green). Imaging shows cytoplasmic and nuclear localization of p70 S6 Kinase (phospho T421 + S424) nuclei (blue) were stained with DAPI.
    F-actin (red) was stained with Alexa Fluor® 594 phalloidin. Images A and B are a composite images. Image C shows results of a competition assay using the p70 S6 Kinase (phospho T421 + S424) peptide.

  • Western blot - Anti-p70 S6 Kinase (phospho T421 + S424) antibody [RP23040018] (ab308099), expandable thumbnail

    Western blot - Anti-p70 S6 Kinase (phospho T421 + S424) antibody [RP23040018] (ab308099)

    Western blot analysis of p70 S6 Kinase (phospho T421 + S424) in whole cell extracts of serum-starved HeLa cells treated with IGF1 (150 ng/mL, 15 min) using ab308099 at a dilution of 1 µg/mL. To confirm specificity, competition was performed by preincubation with the phosphopeptide to inhibit antibody binding (lane 2). Results show a band at ~70 kDa.

    All lanes: Western blot - Anti-p70 S6 Kinase (phospho T421 + S424) antibody [RP23040018] (ab308099) at 1 µg/mL

    Lane 1: serum-starved HeLa cells treated with IGF1

    Lane 2: Preincubation of HeLa cell lysate with phosphopeptide and IGF-1

    Predicted band size: 59 kDa

    Observed band size: 70 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-p70 S6 Kinase (phospho T421 + S424) antibody [RP23040018] (ab308099), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-p70 S6 Kinase (phospho T421 + S424) antibody [RP23040018] (ab308099)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0. 25% Triton X-100 permeabilized 70% confluent log phase U-2 OS cells labeling p70 S6 Kinase (phospho T421 + S424) with ab308099 at 1/200 dilution, followed by Alexa Fluor® 488 Goat anti-Rabbit IgG secondary antibody at 1/400 dilution (Panel a: green). Nuclei (Panel b: blue) were stained with DAPI. F-actin (Panel c: red) was stained with Alexa Fluor® 594 phalloidin. Panel d is a merged image showing cytoplasmic localization respectively. Panel e shows competition with the p70 S6 Kinase (phospho T421 + S424) peptide.
    The images were captured at 20X magnification.

  • Western blot - Anti-p70 S6 Kinase (phospho T421 + S424) antibody [RP23040018] (ab308099), expandable thumbnail

    Western blot - Anti-p70 S6 Kinase (phospho T421 + S424) antibody [RP23040018] (ab308099)

    Western blot analysis of p70 S6 Kinase (phospho T421 + S424) was performed by loading 20 µg of untreated HeLa (lane 1) and HeLa treated with IGF1 (150 ng/mL for 15 minutes) cell lysates (lane 2) using a 4-12% Bis-Tris gel electrophoresis system, and pre-stained protein standard. Proteins were transferred to a nitrocellulose membrane and blocked with 5% skim milk for 1 hour at room temperature. p70 S6 Kinase (phospho T421 + S424) was detected at ~70 kDa using ab308099 at a 1:1000 dilution in 2.5% skim milk at 4°C overnight on a rocking platform. To confirm specificity, competition was performed with the phosphopeptide (10 µg/mL) (lane 3). Detection was performed using an HRP-conjugated Goat anti-Rabbit secondary antibody at a 1:5000 dilution and chemiluminescent detection was performed (ECL)

    All lanes: Western blot - Anti-p70 S6 Kinase (phospho T421 + S424) antibody [RP23040018] (ab308099) at 1/1000 dilution

    Lane 1: Untreated HeLa cell lysate at 20 µg

    Lane 2: HeLa cell lysate treated with IGF1 at 20 µg

    Lane 3: Hela cell lysate treated with IGF-1 and phosphopeptide at 10 µg/mL

    Secondary

    All lanes: HRP-conjugated Goat anti-Rabbit secondary antibody at 1/5000 dilution

    Developed using the ECL technique.

    Predicted band size: 59 kDa

    Observed band size: 70 kDa

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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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