NF-kB p65 ELISA Kit

Catalog No. ABIN8070285

This Colorimetric ELISA kit is designed for the quantitative measurement of Human NF-kB p65.

Quick Overview for NF-kB p65 ELISA Kit (ABIN8070285)

Target: NF-kB p65 (NFkBP65) (Nuclear Factor-kB p65 (NFkBP65))

Binding Specificity: acetylated, total

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Application: ELISA

Sample Type: Cell Lysate, Tissue Lysate

Product Details NF-kB p65 ELISA Kit

Purpose: Measures acetylated NF-KB p65 (RELA) and total NF-KB p65 in human cell and tissue lysates, enabling precise assessment of NF-KB p65 acetylation by ELISA. This assay can be developed for other species (mouse, rat, etc.) upon request. Contact techsupport@raybiotech.com.

Analytical Method: Quantitative

Components:

Size / Qty
• Microplate Coated With Anti-Pan-NF-KB p65 Antibody
• Anti-Acetylation Detection Antibody
• Anti-NF-KB p65 Detection Antibody
• HRP-Conjugated Streptavidin
• 200 μL
• 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
• 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
• Log-log graph paper or computer and software for ELISA data analysis
• Absorbent paper
• Tubes to prepare sample dilutions
• Distilled or deionized water
• Shaker

Application Details

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

Plate: Pre-coated

Protocol:

1. Prepare all reagents and samples as instructed.
2. Add 100 μL 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 Streptavidin 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 NF-kB p65

Target: NF-kB p65 (NFkBP65) (Nuclear Factor-kB p65 (NFkBP65))

Alternative Name: NF-kB P65

Background:

Gene Symbols: RELA, NFKB3

Background: The Acetylated-NF-KB p65 and Total-NF-KB p65 ELISA Kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human acetylated ATP-citrate synthase and total ATP-citrate synthase. An anti-pan NF-KB p65 antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and NF-KB p65 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-acetylation antibody is used to detect acetylated NF-KB p65 or biotinylated anti-NF-KB p65 antibody is used to detect total NF-KB p65 . After washing away unbound antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of acetylated NF-KB p65 or total NF-KB p65 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

Protein Name (Synonyms): Transcription factor p65, Nuclear factor NF-kappa-B p65 subunit, Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3

Gene ID: 5970

UniProt: Q04206

Pathways: NF-kappaB Signaling, RTK Signaling, TCR Signaling, TLR Signaling, Fc-epsilon Receptor Signaling Pathway, Neurotrophin Signaling Pathway, Activation of Innate immune Response, Cellular Response to Molecule of Bacterial Origin, Hepatitis C, Toll-Like Receptors Cascades, S100 Proteins