Human Phospho-PRAS40 (T246) DuoSet IC ELISA

Catalog #: DYC6890-2 Datasheet

Discontinued Product

DYC6890-2 has been discontinued.

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Summary Product Features Kit Content Other Reagents Required Product Datasheets Preparation and Storage Background

Human Phospho-PRAS40 (T246) DuoSet IC ELISA Summary

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Assay Length

4 hours 40 minutes (after plate preparation)

Sample Volume Required

Cell lysates (100 µL)

Assay Range

937.5 - 60,000 pg/mL

Sufficient Materials

Kits available for two, five, or fifteen 96-well plates*

Specificity

Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure phosphorylated in cell lysates. An immobilized capture antibody specific for binds both phosphorylated and unphosphorylated . After washing away unbound material, a biotinylated detection antibody is used to detect only phosphorylated receptor, utilizing a standard HRP format.

Product Features

Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
Development protocols are provided to guide further assay optimization
Assay can be customized to your specific needs
Available in 2, 5, and 15- (96-well) plate pack sizes
Economical alternative to Western blot

Kit Content

Capture Antibody
Conjugated Detection Antibody
Calibrated Immunoassay Standard or Control
Streptavidin-HRP

Other Reagents Required

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na₂HPO₄, 1.5 mM KH₂O₄, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or equivalent

Lysis Buffer*

IC Diluent*

Blocking Buffer*

Substrate Solution: 1:1 mixture of Color Reagent A (H₂O₂) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H₂SO₄ (Catalog # DY994)

Microplates: From Costar EIA Plate (Costar Catalog # 2592) or R&D Systems (Catalog # DY990), or equivalent

Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent

*For the Lysis Buffer, IC Diluent, and Blocking BUffer recommended for a specific DuoSet ELISA Development Kit, please see the product

Product Datasheets

Product Datasheet

Preparation and Storage

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: PRAS40

PRAS40 (40 kDa proline-rich Akt1 substrate), also known as AKT1S1, is a widely expressed component of the cytoplasmic mTOR complex (mTORC1). In response to insulin stimulation, Akt1 phosphorylation of PRAS40 relieves the basal PRAS40-mediated inhibition of mTORC1. Activated mTORC1 is a key effector of the cell survival pathway. PRAS40 phosphorylation by Akt3 or Pim1 similarly exerts an anti-apoptotic effect during tumor progression and neuronal ischemia. Alternate splicing generates an isoform that lacks the N-terminal 130 amino acids.

Long Name:

40 kDa Proline-rich Akt1 Substrate

Entrez Gene IDs:

84335 (Human); 67605 (Mouse); 292887 (Rat)

Alternate Names:

40 kDa; AKT1 substrate 1 (proline-rich); AKT1S1; Lobe; MGC2865; PRAS40; proline-rich AKT1 substrate 1,40 kDa proline-rich AKT substrate