Revision 1

#33229

Store at -20C

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Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, IHC-P, IF-F, IF-IC

Reactivity:
H M R Mk

Sensitivity:
Endogenous

MW (kDa):
70, 85

Source/Isotype:
Rabbit IgG

UniProt ID:
#P23443

Entrez-Gene Id:
6198

Product Usage Information

This product is the carrier free version of product #34475. All data were generated using the same antibody clone in the standard formulation which contains BSA and glycerol.

This formulation is ideal for use with technologies requiring specialized or custom antibody labeling, including fluorophores, metals, lanthanides, and oligonucleotides. It is not recommended for ChIP, ChIP-seq, CUT&RUN or CUT&Tag assays. If you require a carrier free formulation for chromatin profiling, please contact us. Optimal dilutions/concentrations should be determined by the end user.

BSA and Azide Free antibodies are quality control tested by size exclusion chromatography (SEC) to determine antibody integrity.

Formulation

Supplied in 1X PBS (10 mM Na2HPO4, 3 mM KCl, 2 mM KH2PO4, and 140 mM NaCl (pH 7.8)). BSA and Azide Free.

For standard formulation of this product see product #34475

Storage

Store at -20°C. This product will freeze at -20°C so it is recommended to aliquot into single-use vials to avoid multiple freeze/thaw cycles. A slight precipitate may be present and can be dissolved by gently vortexing. This will not interfere with antibody performance.

Specificity/Sensitivity

p70 S6 Kinase (E8K6T) XP® Rabbit mAb recognizes endogenous levels of total p70 S6 kinase protein. This antibody does not cross-react with p70 S6 kinase-2.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu32 of human p70 S6 kinase protein.

Background

p70 S6 kinase is a mitogen activated Ser/Thr protein kinase that is required for cell growth and G1 cell cycle progression (1,2). p70 S6 kinase phosphorylates the S6 protein of the 40S ribosomal subunit and is involved in translational control of 5' oligopyrimidine tract mRNAs (1). A second isoform, p85 S6 kinase, is derived from the same gene and is identical to p70 S6 kinase except for 23 extra residues at the amino terminus, which encode a nuclear localizing signal (1). Both isoforms lie on a mitogen activated signaling pathway downstream of phosphoinositide-3 kinase (PI-3K) and the target of rapamycin, FRAP/mTOR, a pathway distinct from the Ras/MAP kinase cascade (1). The activity of p70 S6 kinase is controlled by multiple phosphorylation events located within the catalytic, linker and pseudosubstrate domains (1). Phosphorylation of Thr229 in the catalytic domain and Thr389 in the linker domain are most critical for kinase function (1). Phosphorylation of Thr389, however, most closely correlates with p70 kinase activity in vivo (3). Prior phosphorylation of Thr389 is required for the action of phosphoinositide 3-dependent protein kinase 1 (PDK1) on Thr229 (4,5). Phosphorylation of this site is stimulated by growth factors such as insulin, EGF and FGF, as well as by serum and some G-protein-coupled receptor ligands, and is blocked by wortmannin, LY294002 (PI-3K inhibitor) and rapamycin (FRAP/mTOR inhibitor) (1,6,7). Ser411, Thr421 and Ser424 lie within a Ser-Pro-rich region located in the pseudosubstrate region (1). Phosphorylation at these sites is thought to activate p70 S6 kinase via relief of pseudosubstrate suppression (1,2). Another LY294002 and rapamycin sensitive phosphorylation site, Ser371, is an in vitro substrate for mTOR and correlates well with the activity of a partially rapamycin resistant mutant p70 S6 kinase (8).

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Applications Key

W: Western Blotting IHC-P: Immunohistochemistry (Paraffin) IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry)

Cross-Reactivity Key

H: Human M: Mouse R: Rat Mk: Monkey

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

SignalStain is a registered trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Western blot analysis of extracts from control mIMCD-3 cells (lane 1) or p70 S6 kinase-1 knockout mIMCD-3 cells (lane 2), and control wild-type immortalized MEF cells (lane3) or p70 S6 Kinase-1/p70 S6 kinase-2 double knockout MEF cells (lane 4), using p70 S6 Kinase (E8K6T) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). The absence of signal in the two knockout models confirms specificity of the antibody for p70 S6 kinase-1. Knockout cells were kindly provided by Dr. Mario Pende, INSERM, Paris, France. Data were generated using the standard formulation of this product.
Western Blotting Image 1: p70 S6 Kinase (E8K6T) Rabbit Monoclonal Antibody (BSA and Azide Free)
Western blot analysis of extracts from various cell lines using p70 S6 Kinase (E8K6T) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). Data were generated using the standard formulation of this product.
Western Blotting Image 2: p70 S6 Kinase (E8K6T) Rabbit Monoclonal Antibody (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded human urothelial carcinoma using p70 S6 Kinase (E8K6T) XP® Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 1: p70 S6 Kinase (E8K6T) Rabbit Monoclonal Antibody (BSA and Azide Free)
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using p70 S6 Kinase (E8K6T) XP® Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 2: p70 S6 Kinase (E8K6T) Rabbit Monoclonal Antibody (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using p70 S6 Kinase (E8K6T) XP® Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 3: p70 S6 Kinase (E8K6T) Rabbit Monoclonal Antibody (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded normal human pancreas using p70 S6 Kinase (E8K6T) XP® Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 4: p70 S6 Kinase (E8K6T) Rabbit Monoclonal Antibody (BSA and Azide Free)
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded normal human parathyroid using p70 S6 Kinase (E8K6T) XP® Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 5: p70 S6 Kinase (E8K6T) Rabbit Monoclonal Antibody (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded mouse testis using p70 S6 Kinase (E8K6T) XP® Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 6: p70 S6 Kinase (E8K6T) Rabbit Monoclonal Antibody (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded mouse kidney using p70 S6 Kinase (E8K6T) XP® Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 7: p70 S6 Kinase (E8K6T) Rabbit Monoclonal Antibody (BSA and Azide Free)
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded mouse thymus using p70 S6 Kinase (E8K6T) XP® Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 8: p70 S6 Kinase (E8K6T) Rabbit Monoclonal Antibody (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded mouse stomach using p70 S6 Kinase (E8K6T) XP® Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 9: p70 S6 Kinase (E8K6T) Rabbit Monoclonal Antibody (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded mouse spleen using p70 S6 Kinase (E8K6T) XP® Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 10: p70 S6 Kinase (E8K6T) Rabbit Monoclonal Antibody (BSA and Azide Free)
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded mouse cerebellum using p70 S6 Kinase (E8K6T) XP® Rabbit mAb. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 11: p70 S6 Kinase (E8K6T) Rabbit Monoclonal Antibody (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma (left), colon carcinoma (middle), or normal breast (right) using p70 S6 Kinase (E8K6T) XP® Rabbit mAb (top) or p70 S6 Kinase Rabbit mAb (bottom). These two antibodies detect unique, non-overlapping epitopes on p70 S6 kinase protein. The similar staining patterns obtained with both antibodies help to confirm the specificity of the staining. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 12: p70 S6 Kinase (E8K6T) Rabbit Monoclonal Antibody (BSA and Azide Free)
Immunohistochemical analysis of paraffin-embedded human small cell carcinoma of the salivary gland using p70 S6 Kinase (E8K6T) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right). Data were generated using the standard formulation of this product.
Immunohistochemistry Image 13: p70 S6 Kinase (E8K6T) Rabbit Monoclonal Antibody (BSA and Azide Free)
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded MEF cell pellet, wild-type (left, positive) or p70 S6 Kinase-1/ p70 S6 Kinase-2 double knockout (right, negative) using p70 S6 Kinase (E8K6T) XP® Rabbit mAb. Cell lines courtesy of Dr. Mario Pende, INSERM, Paris, France. Data were generated using the standard formulation of this product.
Immunohistochemistry Image 14: p70 S6 Kinase (E8K6T) Rabbit Monoclonal Antibody (BSA and Azide Free)
Confocal immunofluorescent analysis of fixed frozen mouse spleen labeled with p70 S6 Kinase (E8K6T)XP(R) Rabbit mAb (left, green) and colabeled with CD45R/B220 (RA3-6B2) Rat mAb (redFluor™ 710 Conjugate) #82984 (center, red). Sections were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (right, blue). Data were generated using the standard formulation of this product.
Immunofluorescence Image 1: p70 S6 Kinase (E8K6T) Rabbit Monoclonal Antibody (BSA and Azide Free)
Confocal immunofluorescent analysis of fixed frozen mouse kidney, labeled with p70 S6 Kinase (E8K6T)XP® Rabbit mAb (left, green) and DyLight™ 554 Phalloidin #13054 (right, red). Sections were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (right, blue). Data were generated using the standard formulation of this product.
Immunofluorescence Image 2: p70 S6 Kinase (E8K6T) Rabbit Monoclonal Antibody (BSA and Azide Free)
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 1

Cell Signaling Technology Logo
Confocal immunofluorescent analysis of MCF7 cells, wild-type (left, positive) or p70 S6 Kinase-1/ p70 S6 Kinase-2 double knockout (right, negative) using p70 S6 Kinase (E6L9T) XP® Rabbit mAb (green) and DAPI #4083 (blue). Cell lines courtesy of Dr. Mario Pende, INSERM, Paris, France. Data were generated using the standard formulation of this product.
Immunofluorescence Image 1: p70 S6 Kinase (E8K6T) Rabbit Monoclonal Antibody (BSA and Azide Free)
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.