AKT1 Kit ELISA

N° du produit ABIN4885966

Ce kit ELISA Colorimetric est destiné à la mesure quantitative de Humain et Souris AKT1.

Aperçu rapide pour AKT1 Kit ELISA (ABIN4885966)

Antigène: AKT1 (V-Akt Murine Thymoma Viral Oncogene Homolog 1 (AKT1))

Épitope: phosphorylated

Reactivité: Humain, Souris

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Application: ELISA

Type d'échantillon: Cell Lysate, Tissue Lysate

Détail du produit AKT1 Kit ELISA

Fonction: Phospho-AKT/GSK beta/mTor ELISA Kit. This assay semi-quantitatively measures phosphorylated AKT/GSK-beta/mTor in lysate samples.

Analytical Method: Semi-Quantitative

Specificité: The antibody pair provided in this kit recognizes phosphorylated AKT (S473), GSK3β (S9), mTOR (S2448), and total AKT, GSK3β, mTOR in Human and Mouse Cell Lysates.

Attributs du produit:

• Simultaneously measure Phosphorylated protein and pan protein in one experiment (for normalization purpose)
• Screen numerous different cell lysates without performing a Western Blot analysis
• Minimal hands-on time, convenient, and non-radioactive material

Ingrédients:

• 1 Microplate (Item A)
• Wash Buffer Concentrate (20x) (Item B)
• Assay Diluent (Item E)
• Detection Antibody Akt (Ser473) (Item C-1)
• Detection Antibody Akt (Item C-2)
• Detection Antibody GSK3β (S9) (Item C-3)
• Detection Antibody GSK3β (Item C-4)
• Detection Antibody mTOR (S2448) (Item C-5)
• Detection Antibody mTOR (Item C-6)
• HRP-conjugated Anti-rabbit IgG (Item D-1)
• HRP-conjugated Streptavidin (Item D-2)
• TMB One-Step Substrate Reagent (Item H)
• Stop Solution (Item I)
• Cell Lysate Buffer (Item J)
• Positive Control 3T3S001-1 (Item K)

,

• 1 Microplate (Item A)
• Wash Buffer Concentrate (20x) (Item B)
• Assay Diluent (Item E)
• Detection Antibody Akt (Ser473) (Item C-1)
• Detection Antibody Akt (Item C-2)
• Detection Antibody GSK3β (S9) (Item C-3)
• Detection Antibody GSK3β (Item C-4)
• Detection Antibody mTOR (S2448) (Item C-5)
• Detection Antibody mTOR (Item C-6)
• HRP-conjugated Anti-rabbit IgG (Item D-1)
• HRP-conjugated Streptavidin (Item D-2)
• TMB One-Step Substrate Reagent (Item H)
• Stop Solution (Item I)
• Cell Lysate Buffer (Item J)
• Positive Control 3T3S001-1 (Item K)

Matériel non inclus:

• Distilled or deionized water
• 100 mL and 1 liter graduated cylinders
• Tubes to prepare sample dilutions
• Protease and Phosphatase inhibitors
• Precision pipettes to deliver 2 μL to 1 mL volumes
• Adjustable 1-25 mL pipettes for reagent preparation
• Benchtop rocker or shaker
• Microplate reader capable of measuring absorbance at 450 nm

Information d'application

Indications d'application: Optimal working dilution should be determined by the investigator.

Plaque: Pre-coated

Protocole:

1. Prepare all reagents and samples as instructed in the manual.
2. Add 100 μL of sample or positive control to each well.
3. Incubate 2.5 h at RT or O/N at 4 °C.
4. Add 100 μL of prepared primary antibody to each well.
5. Incubate 1 h at RT.
6. Add 100 μL of prepared 1X HRP-Streptavidin to each well.
7. Incubate 1 h at RT.
8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 μL of Stop Solution to each well.
11. Read at 450 nm immediately.

Restrictions: For Research Use only

Stockage

Stock: -20 °C

Stockage commentaire: Upon receipt, the kit should be stored at -20°C. Please use within 6 months from the date of shipment. After initial use, Wash Buffer Concentrate (Item B), Assay Diluent (Item E), TMB One-Step Substrate Reagent (Item H), Stop Solution (Item I) and Cell Lysate Buffer (Item J) should be stored at 4°C to avoid repeated freeze-thaw cycles. Return unused wells to the pouch containing desiccant pack, reseal along entire edge, and store at -20°C. Item D, store at 2-8°C for up to one month (store at -20°C for up to 6 months, avoid repeated freeze-thaw cycles). Reconstituted Positive Control (Item K) should be stored at -70°C.

Date de péremption: 6 months

Détail du antigène

Antigène: AKT1 (V-Akt Murine Thymoma Viral Oncogene Homolog 1 (AKT1))

Autre désignation: AKT

Sujet:

Gene Symbols: AKT1, PKB, RAC

Background: AKT Pathway ELISA Sample kit is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in human and mouse cell lysates. By determining phosphorylated AKT, GSK3β and mTOR protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.

This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human and mouse phospho-AKT, total AKT, phospho-GSK3β, total GSK3β, phospho-mTOR and total mTOR. For each target, a capture antibody has been coated onto microwells. Samples are pipetted into the wells and target protein present in a sample is bound to the wells by the immobilized antibody. The wells are washed and a detection antibody is used to detect the captured target protein. After washing away unbound antibody, an HRP-conjugated secondary antibody is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of target protein bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

Protein Name (Synonyms): RAC-alpha serine/threonine-protein kinase (EC 2.7.11.1) (Protein kinase B) (PKB) (Protein kinase B alpha) (PKB alpha) (Proto-oncogene c-Akt) (RAC-PK-alpha)

ID gène: 391

UniProt: P31749

Pathways: Signalisation PI3K-Akt, Signalisation RTK, TCR Signaling, AMPK Signaling, Interferon-gamma Pathway, Signalisation TLR, Fc-epsilon Receptor Signaling Pathway, EGFR Signaling Pathway, Neurotrophin Signaling Pathway, Response to Water Deprivation, Regulation of Actin Filament Polymerization, Carbohydrate Homeostasis, Glycosaminoglycan Metabolic Process, Cellular Glucan Metabolic Process, Regulation of Muscle Cell Differentiation, Cell-Cell Junction Organization, Regulation of Cell Size, Skeletal Muscle Fiber Development, Regulation of Carbohydrate Metabolic Process, Hepatitis C, Protein targeting to Nucleus, CXCR4-mediated Signaling Events, Signaling Events mediated by VEGFR1 and VEGFR2, Negative Regulation of intrinsic apoptotic Signaling, Thromboxane A2 Receptor Signaling, Signaling of Hepatocyte Growth Factor Receptor, Positive Regulation of fat Cell Differentiation, VEGFR1 Specific Signals, VEGF Signaling, L'effet Warburg