PRKAA1 ELISA Kit

Catalog No. ABIN8070559

The Human, Mouse and Rat PRKAA1 ELISA Kit (ABIN8070559) is a Colorimetric ELISA Kit designed to quantify Human, Mouse and Rat PRKAA1.

Quick Overview for PRKAA1 ELISA Kit (ABIN8070559)

Target: PRKAA1 (Protein Kinase, AMP-Activated, alpha 1 Catalytic Subunit (PRKAA1))

Binding Specificity: pThr183, total

Reactivity: Human, Mouse, Rat

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Application: ELISA

Sample Type: Cell Lysate, Tissue Lysate

Product Details PRKAA1 ELISA Kit

Purpose: Phospho-AMPKa1 (T183) and Total AMPKa1 ELISA Kit is a convenient and sensitive assay that can monitor the activation or function of important biological pathways in human, mouse, rat cell lysates.

Analytical Method: Semi-Quantitative

Specificity: The antibody pair provided in this kit recognizes human, mouse and rat AMPK subunit alpha-1 phosphorylated at site Threonine-183.

Components:

• Pre-Coated 96-well Strip Microplate
• Positive Control
• Anti-Phospho Detection Antibody
• Anti-Pan Detection Antibody (*not included in phospho-only version)
• HRP-Conjugated Secondary Antibody
• HRP-Streptavidin (*not included in phospho-only version)
• Wash Buffer
• Assay Diluent
• Lysis Buffer
• TMB One-Step Substrate Reagent
• Stop Solution

Material not included:

• Microplate reader capable of measuring absorbance at 450 nm
• Protease and Phosphatase inhibitors
• Shaker
• Precision pipettes to deliver 2 μL to 1 mL volumes
• Adjustable 1-25 mL pipettes for reagent preparation
• 100 mL and 1 liter graduated cylinders
• Absorbent paper
• Distilled or deionized water
• Log-log graph paper or computer and software for ELISA data analysis
• Tubes to prepare the positive control or sample dilutions

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Plate: Pre-coated

Protocol:

1. Prepare all reagents, samples and positive control as instructed.
2. Add 100 μL positive control or sample to each well. Incubate 2.5 hours at room temperature or overnight at 4 °C with gentle shaking.
3. Add 100 μL prepared detection antibody to each well. Incubate for 1 hour at room temperature with gentle shaking.
4. Add 100 μL prepared HRP-Conjugated solution. Incubate for 1 hour at room temperature with gentle shaking.
5. Add 100 μL TMB One-Step Substrate Reagent to each well. Incubate 30 minutes at room temperature.
6. Add 50 μL 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.

Target Details for PRKAA1

Target: PRKAA1 (Protein Kinase, AMP-Activated, alpha 1 Catalytic Subunit (PRKAA1))

Alternative Name: AMPKa1

Background:

Gene Symbols: PRKAA1, AMPK1

Protein Name (Synonyms): 5'-AMP-activated protein kinase catalytic subunit alpha-1 (AMPK subunit alpha-1), Acetyl-CoA carboxylase kinase (ACACA kinase), Hydroxymethylglutaryl-CoA reductase kinase (HMGCR kinase), Tau-protein kinase PRKAA1

Gene ID: 556210578765248

UniProt: Q13131, Q5EG47, P54645

Pathways: AMPK Signaling, Carbohydrate Homeostasis, Regulation of Carbohydrate Metabolic Process, Warburg Effect