NFkB P52 ELISA Kit

Catalog No. ABIN5690757

Product Details

Target: NFkB P52

Reactivity: Human

Detection Method: Colorimetric

Method Type: DNA-Binding ELISA

Application: ELISA

Purpose: Human NF-κB p52 Transcription Factor Activity Assay. This assay uses a dsDNA coated plate with canonical NF-κB p52 binding sequences to semi-quantitatively detect active NF-κB p52 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 SOX2 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

Target: NFkB P52

Alternative Name: NF-kB P52

Gene ID: 4791

UniProt: Q00653