PPARG ELISA Kit

Catalog No. ABIN5690762

Product Details PPARG ELISA Kit

Target: PPARG (Peroxisome Proliferator-Activated Receptor gamma (PPARG))

Reactivity: Human

Detection Method: Colorimetric

Method Type: DNA-Binding ELISA

Application: ELISA

Purpose:  Human PPAR-gamma Transcription Factor Activity Assay. This assay uses a dsDNA coated plate with canonical PPAR-gamma binding sequences to semi-quantitatively detect active PPAR-gamma in lysates or nuclear extracts. 

Sample Type: Cell Lysate, Nuclear Extract

Analytical Method: Semi-Quantitative

Specificity: The olionucleotide/antibody pair provided in this kit recognizes human PPAR-gamma in whole lysates and nuclear extracts.

Characteristics:

• Specific transcription factor-DNA binding assay
• Perfect alternative to EMSA
• Easy to perform in an ELISA format
• Non-radioactive assay
• High throughput (96 well plate format)
• Assay can be completed within 5 hours

Components:

• 96-well Strip Microplate pre-coated with DNA probes
• DNA Binding Buffer
• Positive Control Sample
• Specific Competitor DNA probe
• Non-specific Competitor DNA probe
• Assay Reagent
• DTT
• Wash Buffer
• Primary Antibody
• HRP-conjugated Secondary Antibody
• TMB One-Step Substrate Reagent
• Stop Solution

Material not included:

• Distilled or deionized water
• 100 mL and 1 liter graduated cylinders
• Tubes to prepare sample dilutions Absorbent paper
• 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

Plate: Pre-coated

Protocol:

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 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 HRP-secondary antibody 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

Handling

Storage: 4 °C

Storage Comment: Upon receipt, the positive control should be removed and stored at -20° or -80°C. The remainder of the kit can be stored for up to 6 months at 2-8°C from the date of shipment. Opened Microplate Wells or reagents may be stored for up to 1 month at 2° to 8°C. Return unused wells to the pouch containing desiccant pack, reseal along entire edge. Note: The kit can be used within one year if the whole kit is stored at -20°C upon receipt. Avoid repeated freeze-thaw cycles.

Expiry Date: 6 months

Target Details for PPARG

Target: PPARG (Peroxisome Proliferator-Activated Receptor gamma (PPARG))

Alternative Name: PPAR-gamma (PPARG Products)

Gene ID: 5468

UniProt: P37231

Pathways: MAPK Signaling, Nuclear Receptor Transcription Pathway, Steroid Hormone Mediated Signaling Pathway, Negative Regulation of Hormone Secretion, Carbohydrate Homeostasis, Regulation of Lipid Metabolism by PPARalpha, Positive Regulation of Endopeptidase Activity, Brown Fat Cell Differentiation, Positive Regulation of fat Cell Differentiation