25% off all ELISA kits

Use code ELISA25-262 for 25% off all ELISAs through November.

25% off all ELISA kits

Use code ELISA25-262 for 25% off all ELISAs through November.

Acetylated NF-KB p65 ELISA Kit

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.

Product Description

Specifications

Size	1 Plate Kit, 2 Plate Kit, 5 Plate Kit
Estimated Lead Time	4-8 weeks
Species	Human
Compatible Sample Types	Tissue Lysates, Cell Lysates
Accession Number	Q04206
Gene Id	5970
Gene Symbols	RELA, NFKB3
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
Solid Support	96-well Microplate
Design Principle	Sandwich-based
Method Of Detection	Colorimetric
Research Area	Post-Translational Modifications, NFkB Signaling, Acetylation, Epigenetics
Modification	Acetylation
Shipping Type	Blue ice
Storage	-20°C

Introduction

The RayBio^® 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.

Product Features

Quantify quickly and easily
Screen numerous different lysates without performing a Western Blot analysis
Minimal hands-on time, convenient, and non-radioactive material

Kit Components

Component	Size / Qty
Microplate Coated With Anti-Pan-NF-KB p65 Antibody	1 plate
Anti-Acetylation Detection Antibody	1 vial
Anti-NF-KB p65 Detection Antibody	1 vial
HRP-Conjugated Streptavidin	200 µL
Wash Buffer	25 mL
Assay Diluent	15 mL
Lysis Buffer	5 mL
TMB One-Step Substrate Reagent	12 mL
Stop Solution	8 mL

Additional Materials Required

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

Assay Procedure Summary

Prepare all reagents and samples as instructed.
Add 100 µL sample to each well. Incubate 2.5 hours at room temperature or overnight at 4°C with gentle shaking.
Add 100 µL prepared detection antibody to each well. Incubate for 1 hour at room temperature with gentle shaking.
Add 100 µL prepared HRP-Conjugated Streptavidin solution. Incubate for 1 hour at room temperature with gentle shaking.
Add 100 µL TMB One-Step Substrate Reagent to each well. Incubate 30 minutes at room temperature.
Add 50 µL Stop Solution to each well. Read at 450 nm immediately.

Storage/Stability

The entire kit may be stored at -20°C for up to 6 months from the date of shipment. Avoid repeated freeze-thaw cycles.

Customization

This assay can be developed for other species (mouse, rat, etc.) upon request. Contact techsupport@raybiotech.com for more information.

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Excel (Sandwich-ELISA-Analysis-Tool.xlsm)

Introduction

Product Features

Quantify quickly and easily
Screen numerous different lysates without performing a Western Blot analysis
Minimal hands-on time, convenient, and non-radioactive material

Kit Components

Component	Size / Qty
Microplate Coated With Anti-Pan-NF-KB p65 Antibody	1 plate
Anti-Acetylation Detection Antibody	1 vial
Anti-NF-KB p65 Detection Antibody	1 vial
HRP-Conjugated Streptavidin	200 µL
Wash Buffer	25 mL
Assay Diluent	15 mL
Lysis Buffer	5 mL
TMB One-Step Substrate Reagent	12 mL
Stop Solution	8 mL

Additional Materials Required

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

Assay Procedure Summary

Prepare all reagents and samples as instructed.
Add 100 µL sample to each well. Incubate 2.5 hours at room temperature or overnight at 4°C with gentle shaking.
Add 100 µL prepared detection antibody to each well. Incubate for 1 hour at room temperature with gentle shaking.
Add 100 µL prepared HRP-Conjugated Streptavidin solution. Incubate for 1 hour at room temperature with gentle shaking.
Add 100 µL TMB One-Step Substrate Reagent to each well. Incubate 30 minutes at room temperature.
Add 50 µL Stop Solution to each well. Read at 450 nm immediately.

Storage/Stability

The entire kit may be stored at -20°C for up to 6 months from the date of shipment. Avoid repeated freeze-thaw cycles.

Customization

This assay can be developed for other species (mouse, rat, etc.) upon request. Contact techsupport@raybiotech.com for more information.

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25% off all ELISA kits

25% off all ELISA kits

Acetylated NF-KB p65 ELISA Kit