Phospho-p70 S6 Kinase (Thr389/Thr412) Antibody

Product:	Phospho-p70 S6 Kinase (Thr389/Thr412) Antibody
Catalog:	AF3228
Description:	Rabbit polyclonal antibody to Phospho-p70 S6 Kinase (Thr389/Thr412)
Application:	WB IHC IF/ICC
Cited expt.:	WB, IHC
Reactivity:	Human, Mouse, Rat, Pig
Prediction:	Pig, Bovine, Horse, Sheep, Rabbit, Dog, Chicken, Xenopus
Mol.Wt.:	70kDa; 59kD(Calculated).
Uniprot:	P23443
RRID:	AB_2834654

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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. By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific.

Size	Price	Inventory
100ul	$280	In stock
200ul	$350	In stock

Lead Time: Same day delivery

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Related Downloads

MSDS

Data Sheet

COA

Protocols

WB Handbook

IHC Protocol

IF/ICC Protocol

Product Info

Source:

Rabbit

Application:

WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user. For optimal experimental results, antibody reuse is not recommended.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:

Human,Mouse,Rat,Pig

Prediction:

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

Clonality:

Polyclonal

Specificity:

Phospho-p70 S6 Kinase (Thr389/Thr412) Antibody detects endogenous levels of p70 S6 Kinase only when phosphorylated at Threonine 389/412.

RRID:

AB_2834654
Cite Format: Affinity Biosciences Cat# AF3228, RRID:AB_2834654.

Conjugate:

Unconjugated.

Purification:

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

Storage:

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.

Alias:

Fold/Unfold

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;

Immunogens

Immunogen:

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

Uniprot:

P23443(KS6B1_HUMAN) >>Visit HPA database.

Gene(ID):

RPS6KB1(6198)

Expression:

P23443 KS6B1_HUMAN:

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:

P23443 KS6B1_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MRRRRRRDGFYPAPDFRDREAEDMAGVFDIDLDQPEDAGSEDELEEGGQLNESMDHGGVGPYELGMEHCEKFEISETSVNRGPEKIRPECFELLRVLGKGGYGKVFQVRKVTGANTGKIFAMKVLKKAMIVRNAKDTAHTKAERNILEEVKHPFIVDLIYAFQTGGKLYLILEYLSGGELFMQLEREGIFMEDTACFYLAEISMALGHLHQKGIIYRDLKPENIMLNHQGHVKLTDFGLCKESIHDGTVTHTFCGTIEYMAPEILMRSGHNRAVDWWSLGALMYDMLTGAPPFTGENRKKTIDKILKCKLNLPPYLTQEARDLLKKLLKRNAASRLGAGPGDAGEVQAHPFFRHINWEELLARKVEPPFKPLLQSEEDVSQFDSKFTRQTPVDSPDDSTLSESANQVFLGFTYVAPSVLESVKEKFSFEPKIRSPRRFIGSPRTPVSPVKFSPGDFWGRGASASTANPQTPVEYPMETSGIEQMDVTMSGEASAPLPIRQPNSGPYKKQAFPMISKRPEHLRMNL

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species

Results

Score

Pig

100

Horse

100

Bovine

100

Sheep

100

Dog

100

Xenopus

100

Chicken

100

Rabbit

100

Zebrafish

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

Research Backgrounds

P23443_KS6B1_HUMAN

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. 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.

PTMs:

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:

Cell junction>Synapse>Synaptosome. Mitochondrion outer membrane. Mitochondrion.
Note: Colocalizes with URI1 at mitochondrion.

Nucleus. Cytoplasm.

Cytoplasm.

Tissue Specificity:

Widely expressed.

Family&Domains:

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.

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 > HIF-1 signaling pathway. (View pathway)

· Environmental Information Processing > Signal transduction > mTOR signaling pathway. (View pathway)

· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway. (View pathway)

· Environmental Information Processing > Signal transduction > AMPK signaling pathway. (View pathway)

· Environmental Information Processing > Signal transduction > TGF-beta signaling pathway. (View pathway)

· Environmental Information Processing > Signal transduction > Apelin signaling pathway. (View pathway)

· Human Diseases > Drug resistance: Antineoplastic > EGFR tyrosine kinase inhibitor resistance.

· Human Diseases > Drug resistance: Antineoplastic > Endocrine resistance.

· Human Diseases > Endocrine and metabolic diseases > Insulin resistance.

· Human Diseases > Infectious diseases: Viral > Human papillomavirus infection.

· Human Diseases > Cancers: Overview > Pathways in cancer. (View pathway)

· Human Diseases > Cancers: Overview > Proteoglycans in cancer.

· Human Diseases > Cancers: Specific types > Colorectal cancer. (View pathway)

· Human Diseases > Cancers: Specific types > Pancreatic cancer. (View pathway)

· Human Diseases > Cancers: Specific types > Acute myeloid leukemia. (View pathway)

· Human Diseases > Cancers: Specific types > Breast cancer. (View pathway)

· Human Diseases > Cancers: Specific types > Hepatocellular carcinoma. (View pathway)

· Human Diseases > Cancers: Specific types > Gastric cancer. (View pathway)

· Human Diseases > Cancers: Overview > Choline metabolism in cancer. (View pathway)

· Organismal Systems > Aging > Longevity regulating pathway. (View pathway)

· Organismal Systems > Aging > Longevity regulating pathway - multiple species. (View pathway)

· Organismal Systems > Immune system > Fc gamma R-mediated phagocytosis. (View pathway)

· Organismal Systems > Endocrine system > Insulin signaling pathway. (View pathway)

References

1). Inhibition of CCT5-mediated asparagine biosynthesis and anti-PD-L1 produce synergistic antitumor effects in colorectal cancer. Acta Pharmaceutica Sinica B, 2025 [IF=14.6]

Application: IHC Species: human Sample:

Figure 7. Pharmacological values of targeting the CCT5/Asn/mTORC1/PD-L1 axis in CRC immunotherapy. (A) IHC staining of CCT5 in tissues from CRC patients receiving PD-L1 therapy. Scale bar = 100 μm. n = 3. (B) Apoptosis of tumor cells after co-culture of PBMCs with Ctrl or CCT5-overexpressing HCT116 cells. n = 3. (C) FACS results of PD-L1 expression in tumor cells after co-culture of PBMCs with Ctrl or CCT5-overexpressing HCT116 cells. n = 3. (D) Functional indices of CD8+ T cells after co-culture of PBMCs with Ctrl or CCT5-overexpressing HCT116 cells. n = 3. (E) The treatment schedule for the monotherapy and combined therapy of ASNase and anti-PD-L1 antibody. (F) Representative images show the effect of ASNase and anti-PD-L1 monotherapy or combination therapy on the growth of subcutaneous tumors. n = 5. (G) The bar charts show the weight of subcutaneous tumors formed by Cct5-manipulated CT26 cells in BALB/c mice under the indicated treatments. n = 5. (H) The line charts illustrate the growth curve of subcutaneous tumors in BALB/c mice after the indicated treatments. The yellow arrow means the start time of the treatment. n = 5. (I) The table displays the TGI of BALB/c mice bearing subcutaneous tumors formed by Cct5-manipulated CT26 cells under the indicated treatments. (J) IHC staining shows the expression of Ki67, p-p70S6K, p-4EBP1, and PD-L1 in subcutaneous tumors. Scale bar = 100 μm. n = 4. See also Fig. S12A. Data are represented as mean ± SD (G, H). ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ns not significant.

2). Endothelial F3-mediated autolysosome and lipid metabolism promote resistance to anti-VEGFA therapy in metastatic colorectal cancer. Autophagy, 2025 (PubMed: 40922383) [IF=14.6]

3). Identification and analysis of microplastic aggregation in CAR-T cells. Journal of hazardous materials, 2024 (PubMed: 39488976) [IF=13.6]

4). Rationally designed rapamycin-encapsulated ZIF-8 nanosystem for overcoming chemotherapy resistance. BIOMATERIALS, 2020 (PubMed: 32841911) [IF=12.8]

Application: WB Species: human Sample: MCF-7/ADR cells

Figure 2.(b) ZIF-8 (10 µg mL−1, 24 h) did not inhibit p70S6K phosphorylation at Thr389 and mTOR phosphorylation at Ser2448 but induced the accumulation of LC3-II, suggesting ZIF-8 elicited mTOR-independent autophagy.

5). Activation of integrated stress response and disordered iron homeostasis upon combined exposure to cadmium and PCB77. JOURNAL OF HAZARDOUS MATERIALS, 2020 (PubMed: 31837937) [IF=12.2]

Application: WB Species: Human Sample: HEL cells

Fig. 5. Disordered iron homeostasis and inhibited mTORC1 activity upon exposure to CdCl2 and PCB77 at low dose. (A) The relative fluorescence intensity of CAeAM for measuring LIP to reflect intracellular iron availability (n = 3–4), and (B) Representative blots of FTH1 protein content to reflect iron storage. Analyses were performed after single or combined exposure to CdCl2 and PCB77 at 1 μM for 48 h. (C) Phosphorylated S6 and total S6 content to reflect mTORC1 activity as measured by Western blot. Ratio of FTH1 to eIF2αP and ratio of pS6 to S6 in the control group were defined as 1. Analyses were performed after single or combined exposure to CdCl2 and PCB77 at 1 μM for 48 h. a- significantly different from the control group. Data were presented in mean ± SE. P < 0.05 was considered statistically significant.

6). A Novel Autophagy Inhibitor p-Hydroxylcinnamaldehyde Suppresses Esophageal Squamous Cell Carcinoma by Targeting LDHA Phosphorylation-Mediated Metabolic Reprogramming. Research (Washington, D.C.), 2026 (PubMed: 41531896) [IF=11.0]

7). Asparagine synthetase modulates glutaminase inhibitor sensitivity through metabolic reprogramming and serves as a prognostic biomarker in hepatocellular carcinoma. Redox biology, 2025 (PubMed: 40779838) [IF=10.7]

8). NCAPD2 inhibits autophagy by regulating Ca2+/CAMKK2/AMPK/mTORC1 pathway and PARP-1/SIRT1 axis to promote colorectal cancer. CANCER LETTERS, 2021 (PubMed: 34229059) [IF=9.1]

Application: WB Species: Human Sample: CRC cells

Fig. 2. NCAPD2 inhibited cell autophagy and disrupted autophagic flux via Ca2+/CAMKK2/AMPK/mTORC1 pathway. (A) Western blot analyses for phosphorylated mTOR (p-mTOR, S2448), phosphorylated p70S6K (p-p70S6K, T389/412), phosphorylated 4E-BP1 (p-4E-BP1, T70) and phosphorylated AKT (p-AKT, S473) in CRCC cells with different treatments as indicated. (B) Western blot of indicated proteins in cells treated with mTORC1 inhibitor Rapamycin (3 nM, 24h). (C) Immunofluorescence staining of LC3II (red) and P62 (red) in CRC cells with different treatments as indicated. Merged images represented overlays of LC3II or P62 and nuclear staining by DAPI (blue). (D) Intracellular Ca2+ levels were analyzed by flow cytometry after staining with the fluorescent probe Fluo-3, AM in CRC cells. (E) Representative Western blot gel documents of phosphorylated CAMKK2(S511), phosphorylated AMPK(T172), phosphorylated mTOR(S2448), Beclin, ATG5, P62, LC3II in CRC cells with different treatments. (F) Western blots of indicated proteins in cells treated with an inhibitor of microsomal Ca2+-ATPase Thapsigargin (1 μM, 6h) and Ca2+ chelator BAPTA-AM (10 μM, 12h) respectively. Results are shown as mean ± s.d, *P < 0.05, **P < 0.01, ***P < 0.001, based on Student’s t-test. . (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

9). Cancer-associated Fibroblast-like Cells Promote Osteosarcoma Metastasis by Upregulation of Phosphoserine Aminotransferase 1 and Activation of the mTOR/S6K Pathway. International journal of biological sciences, 2025 (PubMed: 40612679) [IF=8.2]

Application: WB Species: human Sample: HOS and 143B cells

Figure 6. PSAT1 promotes HOS and 143B migration by activating the mTOR/S6K pathway. (A) mTOR transcriptional levels after siPSAT1 interference in HOS and 143B cells by qPCR. (B) The subcellular positions of PSAT1 and mTOR in HOS and 143B cells by immunofluorescence staining. (C) HOS and 143B cells were treated with 100 μg/mL CHX for 0, 4, 12, and 24 hours in the control and siPSAT1 groups, and the mTOR protein levels were analyzed by western blotting. (D) Western blotting analysis of PSAT1, PI3K, p-PI3K, mTOR, p-mTOR, and p-S6K levels after siPSAT1 in HOS and 143B. (E) PSAT1 overexpression (using pcDNA3.1-PSAT1 overexpression plasmid) in HOS and 143B, and mTOR inhibition (using rapamycin) reducing effector protein S6K phosphorylation. (F) Migration of HOS and 143B after PSAT1 overexpression and mTOR pathway inhibition, analyzed by wound-healing assay, scale bars, 200 µm; the indicated results represent the mean ± SD (n=3), * P < 0.05, ** P < 0.01, *** P < 0.001. (G) Cell migration ability in Transwell assay, scale bars, 100 µm; the indicated results represent the mean ± SD (n=3), * P < 0.05, ** P < 0.01, *** P < 0.001.

10). Inhibition of PTP1B blocks pancreatic cancer progression by targeting the PKM2/AMPK/mTOC1 pathway. Cell Death & Disease, 2019 (PubMed: 31745071) [IF=8.1]

Application: WB Species: Human Sample: pancreatic cancer tissue

Fig. 6 The relationship between PTP1B and AMPK. a PTP1B overexpression resulted in decreased p-AMPK (alpha). b, c The negative correlation between PTP1B and p-AMPKα was showed in pancreatic cancer patient tissue samples (p < 0.001, p value was obtained by a Pearson χ2 test; scale bar, 200 μm and 50 μm). d PTP1B inhibition either by shRNAs or by LXQ46 increased the phosphorylation of PKM2. e, f The inactivated PKM2 resulted in increased phosphorylation of AMPKα and decreased the phosphorylation of PRAS40, causing the inhibition of mTOC1 activity. g PTP1B inhibition caused AMPK activation and decreased p-p70S6K in vivo (scale bar, 200 and 50 μm).

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Phospho-p70 S6 Kinase (Thr389/Thr412) Antibody - #AF3228