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NFkB P52 ELISA Kit

Reactivity: Human Colorimetric DNA-Binding ELISA Cell Lysate, Nuclear Extract
Catalog No. ABIN5690757
  • 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
  • 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
  • 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
    NFkB P52
    Alternative Name
    NF-kB P52
    Gene ID
    4791
    UniProt
    Q00653
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