WB
Western blot analysis of Phospho-p70 S6 Kinase (T389) on different lysates with Rabbit anti-Phospho-p70 S6 Kinase (T389) antibody at 1/1,000 dilution. Lane 1: MCF7 cell lysate, Lane 2: MCF7 starved for 4 hours then treated with 200ng/mL EGF for 15 minutes cell lysate, Lane 3: C6 cell lysate, Lane 4: C6 treated with 100nM Calyculin A for 30 minutes cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 46 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-Phospho-p70 S6 Kinase (T389) antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.ICC/IF
Immunocytochemistry analysis of MCF7 cells treated with or without 20% FBS overnight then add 100nM Calyculin A for 30 minutes labeling Phospho-p70 S6 Kinase (T389) with Rabbit anti-Phospho-p70 S6 Kinase (T389) antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-p70 S6 Kinase (T389) antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.| Product Name | Phospho-p70 S6 Kinase (T389) Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide within human P70 S6 Kinase aa 371-420 / 525. |
| Modification | p-T389 |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ICC/IFIHCWB |
| WB:1:1000-1:2000 IHC:1:1000 ICC:1:100 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | RPS6KB1 |
|---|---|
| Gene Synonyms | S6K PS6K S6K1 STK14A p70-S6K p70 S6KA p70-alpha S6K-beta-1 p70(S6K)-alpha |
| Gene Full Name | ribosomal protein S6 kinase B1 |
| Gene Summary | This gene encodes a member of the ribosomal S6 kinase family of serine/threonine kinases. The encoded protein responds to mTOR (mammalian target of rapamycin) signaling to promote protein synthesis, cell growth, and cell proliferation. Activity of this gene has been associated with human cancer. Alternatively spliced transcript variants have been observed. The use of alternative translation start sites results in isoforms with longer or shorter N-termini which may differ in their subcellular localizations. There are two pseudogenes for this gene on chromosome 17. [provided by RefSeq, Jan 2013] |
| Alternative Names | 70 kDa ribosomal protein S6 kinase 1 antibody p70 alpha antibody p70 S6 kinase alpha antibody p70 S6K antibody p70 S6KA antibody p70(S6K) alpha antibody p70s6kA antibody PS6K antibody Ribosomal protein S6 kinase 70kDa polypeptide 1 antibody ribosomal protein S6 kinase beta 1 antibody 70 kDa ribosomal protein S6 kinase 1 antibody p70 alpha antibody p70 S6 kinase alpha antibody p70 S6K antibody p70 S6KA antibody p70(S6K) alpha antibody p70s6kA antibody PS6K antibody Ribosomal protein S6 kinase 70kDa polypeptide 1 antibody ribosomal protein S6 kinase beta 1 antibody ribosomal protein S6 kinase I antibody Ribosomal protein S6 kinase 70-kD antibody RPS6KB1 antibody S6K antibody S6K1 antibody Serine/threonine kinase 14 alpha antibody Serine/threonine protein kinase 14A antibody STK14A antibody |
| Molecular Weight(MW) | 59kDa(Observed band size: 70/85kDa) |
| Cellular Localization | Synapse, synaptosome, Mitochondrion outer membrane, Mitochondrion. |
WB
Western blot analysis of Phospho-p70 S6 Kinase (T389) on different lysates with Rabbit anti-Phospho-p70 S6 Kinase (T389) antibody at 1/1,000 dilution. Lane 1: MCF7 cell lysate, Lane 2: MCF7 starved for 4 hours then treated with 200ng/mL EGF for 15 minutes cell lysate, Lane 3: C6 cell lysate, Lane 4: C6 treated with 100nM Calyculin A for 30 minutes cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 46 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-Phospho-p70 S6 Kinase (T389) antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ICC/IF
Immunocytochemistry analysis of MCF7 cells treated with or without 20% FBS overnight then add 100nM Calyculin A for 30 minutes labeling Phospho-p70 S6 Kinase (T389) with Rabbit anti-Phospho-p70 S6 Kinase (T389) antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-p70 S6 Kinase (T389) antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594) was used as the secondary antibody at 1/1,000 dilution.| Application Notes | WB:1:1000-1:2000 IHC:1:1000 ICC:1:100 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
