V450 Mouse anti-S6 (pS235/pS236)

BD Phosflow™ V450 Mouse anti-S6 (pS235/pS236)

Name: V450 Mouse anti-S6 (pS235/pS236)
Brand: BD Phosflow™
SKU: 561457

Clone N7-548

(RUO)

Flow cytometric analysis of S6 (pS235/pS236) expression in activated human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were either left untreated (dashed line histogram) or treated with Phorbol 12-Myristate 13-Acetate (PMA; Sigma-Aldrich, Cat. No. P-8139) at 50 nM for 30 minutes (solid line histogram). The cells were then fixed in BD Cytofix™ Fixation Buffer (Cat. No. 554655) at 37°C for 10 minutes, permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050) on ice for 30 minutes, and stained with BD Horizon™ V450 Mouse anti-S6 (pS235/pS236) antibody (Cat. No. 561457). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.

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Product Details

Brand: BD Phosflow™

Alternative Name: 40S ribosomal protein S6; Phosphoprotein NP33; RPS6; RS6

Reactivity: Human (QC Testing)

Isotype: Mouse BALB/c IgG1, κ

Immunogen: Phosphorylated Human ribosomal protein S6 Peptide

Application: Intracellular staining (flow cytometry) (Routinely Tested)

Vol. Per Test: 5 µl

RRID: AB_10643763

Storage Buffer: Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.

Regulatory Status: RUO

Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with BD Horizon™ V450 under optimum conditions, and unreacted BD Horizon™ V450 was removed.

Product Notices

This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
BD Horizon V450 has a maximum absorption of 406 nm and maximum emission of 450 nm. Before staining with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.

Companion Products

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Fixation Buffer RUO

Size 100 mL Cat No. 554655

Perm Buffer III RUO

Size 125 mL Cat No. 558050

561457 Rev. 2

Antibody Details

N7-548

Ribosomal protein S6 (~29 kDa calculated and ~32 kDa observed molecular weights) is a component of the 40S ribosomal subunit and belongs to the S6E family of ribosomal proteins. The S6 ribosomal protein plays a role in regulating the translation of RNAs and thus controlling the growth and proliferation of cells. S6 ribosomal protein phosphorylation, especially at multiple C-terminal serine residues S235, S236, S240, and S244, activates S6. The activated S6 ribosomal protein in turn upregulates the ribosomal translation of RNA species coding for other ribosomal proteins, peptide elongation factors and other proteins involved in cell cycle entry and progression. These phosphorylations are mediated by various kinases (e.g., p70S6K and PKCD) activated through cellular responses to growth factors, cytokines, tumor promoting agents, and mitogens. The S6 ribosomal protein can be dephosphorylated in growth-arrested cells.

The N7-548 monoclonal antibody specifically detects the S6 ribosomal protein phosphorylated at S235 and S236.

The antibody is conjugated to BD Horizon™ V450, which has been developed for use in multicolor flow cytometry experiments and is available exclusively from BD Biosciences. It is excited by the Violet laser Ex max of 406 nm and has an Em Max at 450 nm. Conjugates with BD Horizon™ V450 can be used in place of Pacific Blue™ conjugates.

561457 Rev. 2

Format Details

V450

BD Horizon™ V450 Dye is part of the BD Horizon™ violet family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 405-nm and an emission maximum (Em Max) at 450-nm. BD Horizon™ V450, driven by BD innovation, is designed to be excited by the violet laser (405 nm) and detected using an optical filter centered near 450-nm (e.g., a 450/50-nm bandpass filter). The dye can be excited by the UV (355-nm) laser resulting in cross-laser excitation and spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: V450

Excitation Source: Violet 405 nm

Excitation Max: 405 nm

Emission Max: 450 nm

561457 Rev.2

Citations & References

Development References (9)

Blatt K, Herrmann H, Mirkina I, et al. The PI3-kinase/mTOR-targeting drug NVP-BEZ235 inhibits growth and IgE-dependent activation of human mast cells and basophils. PLoS ONE. 2012; 7(1):e29925. (Clone-specific: Flow cytometry). View Reference
Corradetti MN, Inoki K, Guan KL. The stress-inducted proteins RTP801 and RTP801L are negative regulators of the mammalian target of rapamycin pathway. J Biol Chem. 2005; 280(11):9769-9772. (Biology). View Reference
Gu JJ, Santiago L, Mitchell BS. Synergy between imatinib and mycophenolic acid in inducing apoptosis in cell lines expressing Bcr-Abl. Blood. 2005; 105(8):3270-3277. (Biology). View Reference
Jastrzebski K, Hannan KM, Tchoubrieva EB, Hannan RD, Pearson RB. Coordinate regulation of ribosome biogenesis and function by the ribosomal protein S6 kinase, a key mediator of mTOR function. Growth Factors. 2007; 25(4):209-226. (Biology). View Reference
Kleijn M, Proud CG. The regulation of protein synthesis and translation factors by CD3 and CD28 in human primary T lymphocytes. BMC Biochem. 2002; 3:11. (Biology). View Reference
Lal L, Li Y, Smith J, et al. Activation of the p70 S6 kinase by all-trans-retinoic acid in acute promyelocytic leukemia cells.. Blood. 2005; 105(4):1669-77. (Biology). View Reference
Shah OJ, Ghosh S, Hunter T. Mitotic regulation of ribosomal S6 kinase 1 involves Ser/Thr, Pro phosphorylation of consensus and non-consensus sites by Cdc2. J Biol Chem. 2003; 278(18):16433-16442. (Biology). View Reference
Shah OJ, Hunter T. Critical role of T-loop and H-motif phosphorylation in the regulation of S6 kinase 1 by the tuberous sclerosis complex. J Biol Chem. 2004; 279(20):20816-20823. (Biology). View Reference
van de Laar L, van den Bosch A, Boonstra A, et al. PI3K-PKB hyperactivation augments human plasmacytoid dendritic cell development and function. Blood. 2012; 120(25):4982-4991. (Clone-specific: Flow cytometry). View Reference

View All (9)

561457 Rev. 2

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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.