Thermo Fisher Scientific Phospho-AKT1 (Ser473) Monoclonal Antibody (SDRNR), PE, eBioscience

Applications

Tested Dilution

Publications

Flow Cytometry (Flow)

5 µL (0.06 µg)/test

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Product Specifications

Species Reactivity

Human, Mouse

Published species

Human, Mouse

Host/Isotype

Mouse / IgG2a, kappa

Recommended Isotype Control

Mouse IgG2a kappa Isotype Control (eBM2a), PE, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

SDRNR

Conjugate

PE PE PE

View additional formats

• APC
• Brilliant UV 395
• Brilliant Violet 711
• eFluor 450
• FITC
• PE-Cyanine7
• Request custom conjugation

Excitation/Emission Max

565/576 nm View spectra

Form

Liquid

Concentration

5 µL/Test

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2, with 0.2% BSA

Contains

0.09% sodium azide

Storage conditions

4° C, store in dark, DO NOT FREEZE!

Shipping conditions

Ambient (domestic); Wet ice (international)

RRID

AB_2637098

Product Specific Information

Description: This SDRNR monoclonal antibody recognizes human and mouse AKT (also known as Protein Kinase B (PKB)) when phosphorylated on S473. AKT is a serine/threonine protein kinase that plays a key role in multiple cellular processes including metabolism, proliferation, apoptosis/survival, and migration. There are three homologous isoforms of AKT: AKT1, AKT2, and AKT3. AKT is activated by binding of its pleckstrin homology (PH) domain to membrane phospholipids and by phosphorylation. Phosphorylation of AKT at T308 by PDK1 and at S473 is required for full activation of this kinase. AKT promotes cell survival by inhibiting apoptosis via phosphorylation and inactivation of several targets including Bad, Foxo1, c-Raf, and caspase-9. Deregulation of AKT has been implicated as a major contributing factor in many types of cancer. AKT is negatively regulated by the phosphatase PTEN as well as by the chemical inhibitor LY294002. Specificity of this SDRNR clone was determined by ELISA, flow cytometry, and western blotting.

Applications Reported:This SDRNR antibody has been reported for use in intracellular staining followed by flow cytometric analysis.

Applications Tested: This SDRNR antibody has been pre-titrated and tested by intracellular staining followed by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.06 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.

Staining Protocol: All protocols work well for this monoclonal antibody. Use of Protocol A: Two-step protocol: intracellular (cytoplasmic) proteins allows for the greatest flexibility for detection of surface and intracellular (cytoplasmic) proteins. Use of Protocol B: One-step protocol: intracellular (nuclear) proteins is recommended for staining of transcription factors in conjunction with surface and phosphorylated intracellular (cytoplasmic) proteins. Protocol C: Two-step protocol: Fixation/Methanol allows for the greatest discrimination of phospho-specific signaling between unstimulated and stimulated samples, but with limitations on the ability to stain specific surface proteins (refer to Clone Performance Following Fixation/Permeabilization located in the BestProtocols Section under the Resources tab online). All Protocols can be found in the Flow Cytometry Protocols: Staining Intracellular Antigens for Flow Cytometry Protocol located in the BestProtocols® Section under the Resources tab online.

Excitation: 488-561 nm; Emission: 578 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

AKT1 (PKB alpha) is a serine/threonine kinase that regulates cell survival. The activated enzyme inhibits apoptosis and stimulates cell cycle progression by phosphorylating numerous targets in various cell types, including cancer cells. This protein kinase is activated by insulin, PI3K, IGF1 and various other growth and survival factors. Akt promotes cell survival by inhibiting apoptosis through phosphorylation and inactivation of several targets, including forkhead transcription factors, and caspase-9. The AKT pathway is a major target for cancer drug discovery.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.