Recombinant Anti-S6K1 antibody [E175] (ab32359)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E175] to S6K1
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-S6K1 antibody [E175]
See all S6K1 primary antibodies -
Description
Rabbit monoclonal [E175] to S6K1 -
Host species
Rabbit -
Specificity
This antibody may detect both phosphorylated and non-phosphorylated forms of p70 S6 Kinase.
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Tested applications
Suitable for: WB, IHC-Pmore details
Unsuitable for: Flow Cyt or ICC/IF -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Rat, Cow -
Immunogen
Synthetic peptide within Human S6K1. The exact sequence is proprietary.
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Positive control
- Jurkat cell lysate; human breast carcinoma WB: C-Myc/DDK tagged human S6K1 recombinant protein, C-Myc/DDK tagged human S6K2 recombinant protein.
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General notes
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
E175 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab32359 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB | (1) |
1/1000 - 1/5000. Detects a band of approximately 70 kDa (predicted molecular weight: 59 kDa).
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IHC-P | (2) |
1/50. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Notes |
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WB
1/1000 - 1/5000. Detects a band of approximately 70 kDa (predicted molecular weight: 59 kDa). |
IHC-P
1/50. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Target
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Function
Acts to integrate nutrient and growth factor signals in regulation of protein synthesis, cell proliferation, cell growth, cell cycle progression and cell survival. Downstream effector of the mTOR signaling pathway. Phosphorylates specifically ribosomal protein S6 in response to insulin or several classes of mitogens. During translation initiation, the inactive form associatess with the eIF-3 complex under conditions of nutrient depletion. Mitogenic stimulation leads to phosphorylation and dissociation from the eIF-3 complex and the free activated form can phosphorylate other translational targets including EIF4B. Promotes protein synthesis by phosphorylating PDCD4 at 'Ser-67' and targeting it for degradation. Phosphorylates RICTOR leading to regulation of mammalian target of rapamycin complex 2 (mTORC2) signaling; probably phosphorylates RICTOR at 'Thr-1135'. Phosphorylates IRS1 at multiple serine residues coupled with insulin resistance; probably phosphorylates IRS1 at 'Ser-270'. Required for TNF-alpha induced IRS-1 degradation. Phosphorylates EEF2K in response to IGF1 and inhibits EEF2K activity. Phosphorylates BAD at 'Ser-99' in response to IGF1 leading to BAD inactivation and inhibition of BAD-induced apoptosis. Phosphorylates mitochondrial RMP leading to dissociation of a RMP:PPP1CC complex; probably phosphorylates RMP at 'Ser-99'. The free mitochondrial PPP1CC can dephosphorylate RPS6KB1 at Thr-412 which is proposed to be a negative feed back mechanism for the RPS6KB1 antiapoptotic function. Phosphorylates GSK3B at 'Ser-9' under conditions leading to loss of the TSC1-TSC2 complex. Phosphorylates POLDIP3. -
Tissue specificity
Widely expressed. -
Sequence similarities
Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. S6 kinase subfamily.
Contains 1 AGC-kinase C-terminal domain.
Contains 1 protein kinase domain. -
Domain
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. -
Post-translational
modificationsPhosphorylation at Thr-412 is regulated by mTORC1. The phosphorylation at this site is maintained by an agonist-dependent autophosphorylation mechanism. -
Cellular localization
Cytoplasm; Nucleus. Cytoplasm and Cell junction > synapse > synaptosome. Mitochondrion outer membrane. - Information by UniProt
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Database links
- Entrez Gene: 404181 Cow
- Entrez Gene: 6198 Human
- Entrez Gene: 72508 Mouse
- Entrez Gene: 83840 Rat
- Omim: 608938 Human
- SwissProt: P23443 Human
- SwissProt: Q8BSK8 Mouse
- SwissProt: P67999 Rat
see all -
Alternative names
- 70 kDa ribosomal protein S6 kinase 1 antibody
- KS6B1_HUMAN antibody
- p70 alpha antibody
see all
Images
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All lanes : Anti-S6K1 antibody [E175] (ab32359) at 1/1000 dilution
Lane 1 : C-Myc/DDK tagged human S6K1 recombinant protein 10ng
Lane 2 : C-Myc/DDK tagged human S6K2 recombinant protein 10ng
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 59 kDa
Observed band size: 60,75 kDa why is the actual band size different from the predicted?
Exposure time: 10 secondsBlocking and dilution buffer and concentration: 5% NFDM/TBST.
ab205606 was also used at 1/1000 dilution to detect C-Myc/DDK tagged human S6K1 and S6K2 recombinant proteins shown on the right panel.
ab32539 showed cross reactivity with S6K2 in recombinant protein testing. S6K1 and S6K2 have different observed band size in endogenous and recombinant protein tests.
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: S6K1 knockout HAP1 cell lysate (20 µg)
Lane 3: MCF7 cell lysate (20 µg)
Lane 4: HEK293 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab32359 observed at 68 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab32359 was shown to recognize S6K1 when S6K1 knockout samples were used, along with additional cross-reactive bands. Wild-type and S6K1 knockout samples were subjected to SDS-PAGE. ab32359 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging. -
Paraffin-embedded human breast carcinoma
ab32359 at 1/50 dilutionPerform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Anti-S6K1 antibody [E175] (ab32359) + HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Predicted band size: 59 kDa
Datasheets and documents
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SDS download
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Datasheet download
References (61)
ab32359 has been referenced in 61 publications.
- Wang Y et al. The synergistic antitumor effect of IL-6 neutralization with NVP-BEZ235 in hepatocellular carcinoma. Cell Death Dis 13:146 (2022). PubMed: 35165269
- Zheng H et al. Dysregulated pseudogene BNIP3P1 inhibited cell proliferation and promoted cell apoptosis in preeclampsia by acting as a competing endogenous RNA for BNIP3. Environ Toxicol 37:971-982 (2022). PubMed: 35187800
- Xu MN et al. Enhanced Activation of mTOR Signaling Pathway Was Found in the Hypertrophic and Nodular Lesions of Port Wine Stains. Clin Cosmet Investig Dermatol 15:643-651 (2022). PubMed: 35444442
- Ouyang H et al. Impaired expression of BCAT1 relates to muscle atrophy of mouse model of sarcopenia. BMC Musculoskelet Disord 23:450 (2022). PubMed: 35562710
- Dong X et al. Effect of serotonin on the cell viability of the bovine mammary alveolar cell-T (MAC-T) cell line. J Anim Sci Technol 64:922-936 (2022). PubMed: 36287778