AKT1 ELISA Kit

Produktnummer ABIN4881275

Produktdetails AKT1 ELISA Kit

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

Bindungsspezifität: pSer473

Reaktivität: Human, Maus, Ratte

Nachweismethode: Colorimetric

Methodentyp: Cell ELISA

Applikation: ELISA

Verwendungszweck: Cell-Based Human/Mouse/Rat AKT (S473) Phosphorylation ELISA Kit. Suitable for adherent whole cell lines.

Proben: Cell Culture Cells

Analytische Methode: Semi-Quantitative

Spezifität: The antibodies provided in this kit recognizes human/mouse/rat AKT phosphorylated at Serine-473 and total AKT for comparison.

Produktmerkmale:

• Site and signal pathway-specific
• In vitro detection of adherent cell culture
• No sample lysis needed
• Compatible with a standard ELISA plate reader
• Faster results than with ELISA
• Adaptable for high-throughput screening and drug discovery

Bestandteile:

• uncoated 96-well Microplate
• Wash Buffer A
• Wash Buffer B
• Fixing Solution
• Quenching Buffer
• Blocking Buffer
• Anti-phospho antibody
• Anti-pan antibody
• HRP-Conjugated Secondary Antibody
• TMB One-Step Substrate
• Stop Solution

Benötigtes Material:

• 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

Anwendungsinformationen

Probenmenge: 100 μL

Plattentyp: Uncoated

Protokoll:

1. Seed 10,000-30,000 cells into each well and incubate overnight.
2. Apply various treatment, inhibitors or activators according to manufacture's instructions.
3. Add 100 μL of Fixing Solution into each well and incubate for 20 min at RT with shaking.
4. Add 200 μL of prepared 1X Quenching Buffer and incubate 20 min at RT.
5. Add 200 μL of Blocking Solution and incubate for 1 h at 37 °C.
6. Add 50 μL of 1X anti-phospho-protein specific antibody or anti-pan-protein specific antibody to each well and incubate for 2 h at RT.
7. Add 50 μL of prepared 1X HRP-Anti-Rabbit or Mouse IgG and incubate for 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.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Lagerung: -20 °C

Informationen zur Lagerung: The entire kit may be stored at -20°C for up to 6 months from the date of shipment. Avoid repeated freeze-thaw cycles.

Haltbarkeit: 6 months

Details zu AKT1

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

Andere Bezeichnung: Akt (AKT1 Produkte)

Gen-ID: 207

UniProt: P31749

Pathways: PI3K-Akt Signalweg, RTK Signalweg, T-Zell Rezeptor Signalweg, AMPK Signaling, Interferon-gamma Pathway, TLR Signalweg, Fc-epsilon Rezeptor Signalübertragung, EGFR Signaling Pathway, Neurotrophin Signalübertragung, 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, Warburg Effekt