Control Lysate
(MCF-7 cells were grown in a T175 flask at 37 degree C, 5% CO2, in EMEM with 10% FBS,10% non-essential amino-acidsand cultured to 90% confluence. The medium was removed and cells were stimulated with 30 uM Insulin for 60 min at room temperature. The medium was removed and cells were lysed with 4 mL of 1X Lysis Buffer #3 supplemented with 1 mM NaF and 2 mM Na3VO4, on an orbital shaker (400 rpm) for 30 min at room temperature. Soluble supernatants were collected after a 10 min centrifugation and tested using the two-plate assay protocol.)
Human p70 S6K Assay Kit | RPS6KB1 assay kit
Phospho-p70 S6K (T389) TR-FRET Cellular Assay Kit
Acc-phospho-P70S6 (20 uL)
P70S6 control lysate (100 uL)
5X Lysis Buffer #3 (1 mL)
10X TR-FRET Detection buffer (50 uL)
Control Lysate
(MCF-7 cells were grown in a T175 flask at 37 degree C, 5% CO2, in EMEM with 10% FBS,10% non-essential amino-acidsand cultured to 90% confluence. The medium was removed and cells were stimulated with 30 uM Insulin for 60 min at room temperature. The medium was removed and cells were lysed with 4 mL of 1X Lysis Buffer #3 supplemented with 1 mM NaF and 2 mM Na3VO4, on an orbital shaker (400 rpm) for 30 min at room temperature. Soluble supernatants were collected after a 10 min centrifugation and tested using the two-plate assay protocol.)
Control Lysate
(MCF-7 cells were grown in a T175 flask at 37 degree C, 5% CO2, in EMEM with 10% FBS,10% non-essential amino-acidsand cultured to 90% confluence. The medium was removed and cells were stimulated with 30 uM Insulin for 60 min at room temperature. The medium was removed and cells were lysed with 4 mL of 1X Lysis Buffer #3 supplemented with 1 mM NaF and 2 mM Na3VO4, on an orbital shaker (400 rpm) for 30 min at room temperature. Soluble supernatants were collected after a 10 min centrifugation and tested using the two-plate assay protocol.)
Testing Data
(MCF-7 cells were seeded at 75,000 cells/well and cultured overnight at 37 degree C, 5% CO2, in EMEM with 10% FBS and 10% non-essential amino-acids. Cells were then treated at room temperature with either Insulin for 60 min(stimulation) or LY294002 for 60 min then 1 uM Insulin(inhibition).The media was removed from the wells, and cells were lysed with 50 uL/well of 1X Lysis Buffer #3 containing 1 mM NaF and 2 mM Na3VO4, on an orbital shaker (400 rpm) for 30 min at room temperature. The lysates were then analyzed for phospho-p70 S6K (T389)using the two-plate assay protocol. The TR-FRET signal was recorded at 665 and 615 nm (EnVision lamp excitation) after an18 h incubation period at room temperature.)
Testing Data
(MCF-7 cells were seeded at 75,000 cells/well and cultured overnight at 37 degree C, 5% CO2, in EMEM with 10% FBS and 10% non-essential amino-acids. Cells were then treated at room temperature with either Insulin for 60 min(stimulation) or LY294002 for 60 min then 1 uM Insulin(inhibition).The media was removed from the wells, and cells were lysed with 50 uL/well of 1X Lysis Buffer #3 containing 1 mM NaF and 2 mM Na3VO4, on an orbital shaker (400 rpm) for 30 min at room temperature. The lysates were then analyzed for phospho-p70 S6K (T389)using the two-plate assay protocol. The TR-FRET signal was recorded at 665 and 615 nm (EnVision lamp excitation) after an18 h incubation period at room temperature.)
A superior alternative to ELISA
Features :: Benefits
• TR-FRET technology :: Proven, sensitive and robust assay technology
• Homogeneous, add-and-read assay :: No wash, no separation steps
• < 1 hour bench time :: Faster, easier than ELISA
• Adherent or suspension cells :: Higher flexibility
• Validated assays :: Results you can trust
• Small sample volume (15uL) :: Save precious samples
• Compatible with low-volume 96-well and 384-well plates : Higher throughput
• Signal stable over time :: Flexibility to read plate
• Kits available in 100 and 500 assay points :: Cost-effective and flexible
• Kits available for phospho, total and phospho+total protein :: Allows multiple protein measurements in one plate from one sample
NCBI and Uniprot Product Information
NCBI Description
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]
Uniprot Description
p70S6K: an AGC kinase of the RSK family that is required for cell growth and G1 cell cycle progression. Is phosphorylated and activated by mTOR in mitogenic pathways downstream of phosphoinositide 3 kinase (PI3K). Phosphorylates the S6 protein of the 40S ribosomal subunit and is involved in translational control of 5' oligopyrimidine tract mRNAs. Activity is controlled by multiple phosphorylation events located within the catalytic, linker and pseudosubstrate domains. Mouse knockout shows symptoms of insulin resistance, and increased insulin senstivity, resulting in protection against diet-induced obesity. Protein expression and activation upregulated in colon adenocarcinoma cell lines. Increased expression in breast cancer correlated with poor survival. Selectively amplified and overexpressed within the 17q23 breast cancer amplicon. Two isoforms produced by alternative initiation have been reported.
Protein type: EC 2.7.11.1; Protein kinase, Ser/Thr (non-receptor); Protein kinase, AGC; Kinase, protein; Translation; AGC group; RSK family; p70 subfamily
Chromosomal Location of Human Ortholog: 17q23.1
Research Articles on RPS6KB1
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Product Notes
The Human RPS6KB1 rps6kb1 (Catalog #AAA298047) is an Assay Kit and is intended for research purposes only. The product is available for immediate purchase. The AAA298047 Assay Kit recognizes Human RPS6KB1. AAA Biotech's p70 S6K can be used in a range of immunoassay formats including, but not limited to, Cell-Based Assays. Researchers should empirically determine the suitability of the RPS6KB1 rps6kb1 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "p70 S6K, Assay Kit" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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