CoraLite®555-conjugated Phospho-AKT1 (Ser473) Recombinant antibody

Phospho-AKT1 (Ser473) Recombinant Antibody for IF/ICC, FC (Intra)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse

Applications

IF/ICC, FC (Intra)

Conjugate

CoraLite®555 Fluorescent Dye

CloneNo.

2M10

Cat no : CL555-80462

Synonyms

AKT pan, AKT 1, AKT (pan), AKT, AKT1

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Validation Data Gallery

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  IF Staining of HeLa using CL555-80462

  Immunofluorescent analysis of (4% PFA) fixed Calyculin A treated HeLa cells using CoraLite®555 Phospho-AKT1 (Ser473) antibody (CL555-80462, Clone: 2M10 ) at dilution of 1:200.

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  FC experiment of NIH/3T3 using CL555-80462

  1X10^6 NIH/3T3 cells untreated (dashed line) or treated with Calyculin A (red) were intracellularly stained with 0.25 ug CoraLite®555 Anti-Human Phospho-AKT1 (Ser473) (CL555-80462, Clone:2M10), or 0.25 ug Control Antibody (Blue). Cells were fixed with 4% PFA and permeabilized with 90% MeOH.

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Tested Applications

• Positive IF/ICC detected in: Calyculin A treated HeLa cells
• Positive FC (Intra) detected in: Calyculin A treated NIH/3T3 cells

Recommended dilution

• Immunofluorescence (IF)/ICC: IF/ICC : 1:50-1:500
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.25 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

CL555-80462 targets Phospho-AKT1 (Ser473) in IF/ICC, FC (Intra) applications and shows reactivity with human, mouse samples.

• Tested Reactivity: human, mouse
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: Peptide
• Full Name: v-akt murine thymoma viral oncogene homolog 1
• Observed Molecular Weight: 56-62 kDa
• GenBank Accession Number: NM_005163
• Gene Symbol: AKT1
• Gene ID (NCBI): 207
• RRID: AB_2919710
• Conjugate: CoraLite®555 Fluorescent Dye
• Excitation/Emission Maxima Wavelengths: 557 nm / 570 nm
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 50% Glycerol, 0.05% Proclin300, 0.5% BSA, pH 7.3.
• Storage Conditions: Store at -20°C. Avoid exposure to light. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage.

Background Information

AKT is a serine/threonine kinase and it participates in the key role of the PI3K signaling pathway. Phosphatidylinositol-3 kinase (PI3K) is the key regulator of AKT activation. The recruitment of inactive AKT protein to PIP3-rich areas of the plasma membrane results in a conformational change that exposes the activation loop of AKT. AKT's activating kinase, phosphoinositide-dependent protein kinase (PDK1) , is also recruited to PIP3 microdomains. PDK1 phosphorylates AKT on threonine 308 (Thr308) of the exposed activation loop, activating AKT and leading to a second phosphorylation of AKT at serine 473 (Ser473) by a kinase presumed to be mTORC2 that further potentiates kinase activity. Active AKT will phosphorylate various downstream protein targets that control cell growth and translational control and act to suppress apoptosis. (PMID: 31594388, PMID: 30808672). 80462-1-RR specifically recognizes AKT1 phosphorylated at Ser473.