MTOR ELISA Kit

Produktnummer ABIN2748394

Dieses Human MTOR ELISA-Kit ist ein Colorimetric ELISA-Kit, das dafür entwickelt wurde, Human MTOR in Cell Culture Cells zu quantifizieren.

Kurzübersicht für MTOR ELISA Kit (ABIN2748394)

Target: MTOR (mTOR) (Mechanistic Target of Rapamycin (serine/threonine Kinase) (mTOR))

Bindungsspezifität: pSer2448

Reaktivität: Human

Nachweismethode: Colorimetric

Methodentyp: Cell ELISA

Applikation: ELISA

Proben: Cell Culture Cells

Produktdetails MTOR ELISA Kit

Verwendungszweck: Cell-Based Human mTOR (S2448) Phosphorylation ELISA Kit. Suitable for adherent whole cell lines.

Analytische Methode: Semi-Quantitative

Spezifität: The antibodies provided in this kit recognizes human and mouse mTOR phosphorylated at Serine-2448 and total mTOR 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.

Testdurchführung:

Seed 10,000-30,000 cells into each well and incubate overnight.
Apply various treatment, inhibitors or activators according to manufacture's instructions.
Add 100 μL of Fixing Solution into each well and incubate for 20 min at RT with shaking.
Add 200 μL of prepared 1X Quenching Buffer and incubate 20 min at RT.
Add 200 μL of Blocking Solution and incubate for 1 h at 37 °C.
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.
Add 50 μL of prepared 1X HRP-Anti-Rabbit or Mouse IgG and incubate for 1 h at RT.
Add 100 μL of TMB One-Step Substrate Reagent to each well.
Incubate 30 min at RT.
Add 50 μL of Stop Solution to each well.
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 MTOR

Target: MTOR (mTOR) (Mechanistic Target of Rapamycin (serine/threonine Kinase) (mTOR))

Andere Bezeichnung: mTOR

Hintergrund: Mechanistic target of rapamycin, mammalian target of rapamycin (mTOR), FK506-binding protein 12-rapamycin-associated protein 1 (FRAP1)

Gen-ID: 2475

UniProt: P42345

Pathways: PI3K-Akt Signalweg, RTK Signalweg, AMPK Signaling, Interferon-gamma Pathway, Fc-epsilon Rezeptor Signalübertragung, EGFR Signaling Pathway, Neurotrophin Signalübertragung, Regulation of Actin Filament Polymerization, Regulation of Muscle Cell Differentiation, Regulation of Cell Size, Skeletal Muscle Fiber Development, Regulation of Carbohydrate Metabolic Process, Autophagie, CXCR4-mediated Signaling Events, BCR Signaling, Warburg Effekt