PE Mouse anti-4EBP1 (pT36/pT45)

BD Phosflow™ PE Mouse anti-4EBP1 (pT36/pT45)

Name: PE Mouse anti-4EBP1 (pT36/pT45)
Brand: PE Mouse anti-4EBP1 (pT36/pT45)
SKU: 560285

Clone M31-16 (RUO)

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Product Details

Brand: BD Phosflow™

Alternative Name: 4E-BP1. EIF4EBP1, P/OKCL.6, PHAS-I, PHAS-1

Reactivity: Human (QC Testing), Mouse, Rat (Predicted)

Isotype: Mouse BALB/c IgG1, κ

Immunogen: Phosphorylated Human 4EBP1 (pT45) Peptide

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

Vol. Per Test: 20 µl

RRID: AB_1645498

Storage Buffer: Aqueous buffered solution containing BSA 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 R-PE under optimum conditions, and unconjugated antibody and free PE were removed.

Recommended Assay Procedures

Either BD Cytofix™ fixation buffer or BD Phosflow™ Fix Buffer I may be used for cell fixation. Any of the three BD Phosflow™ permeabilization buffers may be used.

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).
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
Source of all serum proteins is from USDA inspected abattoirs located in the United States.

Companion Products

Fixation Buffer RUO

Size 100 ML Cat No. 554655

Fix Buffer I RUO

Size 250 ML Cat No. 557870

Perm/Wash Buffer I RUO

Size 125 ML Cat No. 557885

Perm Buffer II RUO

Size 125 ML Cat No. 558052

Perm Buffer III RUO

Size 125 ML Cat No. 558050

560285 Rev. 2

Antibody Details

M31-16

The eukaryotic translation initiation factor 4E-Binding Protein 1 (4EBP1) is a phosphorylated heat- and acid-stable protein (PHAS-I or PHAS-1), and it is regulated by insulin. It is a member of the eIF4E-Binding Protein Family, which also includes the proteins 4EBP2 and 4EBP3. 4EBP1 binds with eukaryotic translation Initiation Factor 4E (eIF4E), which prevents its assembly into the eIF4E complex and inhibits cap-dependent translation. When 4EBP1 is phosphorylated, this binding is disrupted, allowing cap-dependent translation to be activated. Phosphorylation of 4EBP1 is required for protein synthesis, and it mediates the regulation of protein translation by stimuli that signal through the phosphoinositide 3 (PI3) kinase pathway. We found that threonines 36 and 45 (T36/T45) are phosphorylated in resting human peripheral blood mononuclear cells. PI3 kinase inhibitors, such as LY294002 down-regulate the phosphorylation level of 4EBP1 (pT36/pT45).

The M31-16 monoclonal antibody recognizes the phosphorylated T36 and T45 of activated human 4EBP1. The orthologous phosphorylation sites in mouse and rat 4EBP1 are T35 and T44.

560285 Rev. 2

Format Details

R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: PE

Excitation Max: 496 nm, 566 nm

Emission Max: 576 nm

560285 Rev.2

Citations & References

View product citations for antibody "560285" on CiteAb

Development References (2)

Gingras AC, Raught B, Gygi SP, et al. Hierarchical phosphorylation of the translation inhibitor 4E-BP1. Genes Dev. 2001; 15(21):2852-2864. (Biology). View Reference
Hay N, Sonenberg N. Upstream and downstream of mTOR. Genes Dev. 2004; 18:1926-1945. (Biology).

560285 Rev. 2

Please refer to Support Documents for Quality Certificates

Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described

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.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product as described.

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.