MTOR ELISA Kit

Catalog No. ABIN2748394

Product Details MTOR ELISA Kit

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

Binding Specificity: pSer2448

Reactivity: Human

Detection Method: Colorimetric

Method Type: Cell ELISA

Application: ELISA

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

Sample Type: Cell Culture Cells

Analytical Method: Semi-Quantitative

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

Characteristics:

• 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

Components:

• 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

Material not included:

• 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

Application Details

Sample Volume: 100 μL

Plate: Uncoated

Protocol:

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.

Assay Procedure:

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.

Restrictions: For Research Use only

Handling

Storage: -20 °C

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

Expiry Date: 6 months

Target Details for MTOR

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

Alternative Name: mTOR (mTOR Products)

Synonyms: FRAP1 ELISA Kit, FRAP ELISA Kit, FRAP2 ELISA Kit, RAFT1 ELISA Kit, RAPT1 ELISA Kit, 2610315D21Rik ELISA Kit, AI327068 ELISA Kit, Frap1 ELISA Kit, flat ELISA Kit, frap1 ELISA Kit, tor ELISA Kit, wu:fc22h08 ELISA Kit, mechanistic target of rapamycin kinase ELISA Kit, mechanistic target of rapamycin ELISA Kit, mechanistic target of rapamycin (serine/threonine kinase) ELISA Kit, MTOR ELISA Kit, Mtor ELISA Kit, mtor ELISA Kit

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

Gene ID: 2475

UniProt: P42345

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