Human/Mouse/Rat Phospho-NF-KB p65 (S536) and Total NF-KB p65 ELISA Kit

RayBiotech社のHuman/Mouse/Rat Phospho-NF-KB p65 (S536) and Total NF-KB p65 ELISA Kitです。
※本製品は研究用です。研究用以外には使用できません。

■Introduction

RayBio® Phospho- NF-κB p65(S536) and Total NF-κB p65 ELISA kit is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in human, mouse and rat cell lysates. By determining phosphorylated NF-κB p65 protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.

This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human, mouse and rat phospho- NF-κB p65 and total NF-κB p65. An anti-pan NF-κB p65 antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and NF-κB p65 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and rabbit anti- NF-κB p65 (S536) antibody is used to detect phosphorylated NF-κB p65 or biotinylated anti-NF-κB p65 antibody is used to detect pan NF-κB p65. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG or 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 NF-κB p65(S536) or pan NF-κB 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

■Specificity

The antibody pair provided in this kit recognizes human NF-KB p65 phosphorylated at site Serine-536 and Total NF-KB p65

■Target Protein Information

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

Accession Number: Q04206

Gene Names: RELA NFKB3

Gene ID: 5970

■Species Detected

Human

■Application Notes

Kit Components

Other Materials Required

Protocol Outline

■Typical Data

Positive Control

HeLa cells were treated with TNFα and Calyculin A at 37°C for 5 min. Solubilize cells at 4 x 10^7 cells/ml in Cell Lysate Buffer. Serial dilutions of lysates were analyzed in this ELISA. Please see step 3 of Part VI Reagent Preparation for detail.

Calyculin A Stimulation of HeLa Cell Line

HeLa cells were treated or untreated with TNFα and 100nM Calyculin A for 5 min. Cell lysates were analyzed using this phosphoELISA and Western Blot.

■Storage/Stability

Upon receipt, the kit should be stored at –20°C. Please use within 6 months from the date of shipment. After initial use, Wash Buffer Concentrate (Item B), Assay Diluent (Item E), TMB One-Step Substrate Reagent (Item H), Stop Solution (Item I) and Cell Lysate Buffer (Item J) should be stored at 4°C to avoid repeated freeze-thaw cycles. Return unused wells to the pouch containing desiccant pack, reseal along entire edge, and store at –20°C. Item D, store at 2-8°C for up to one month (store at -20°C for up to 6 months, avoid repeated freeze-thaw cycles). Reconstituted Positive Control (Item K) should be stored at -70°C.

■ Antigen Information

Gene Symbol:

■ Product Specifications

Size: 1, 2, or 5 x 96-Well Strip Microplate Kit

■ Array/ELISA Format

Pathway:

Species Detected:

Solid Support: 96-well Microplate

Design Principle:

Quantitative/Semi-Quantitative: Semi-Quantitative

Method of Detection: Colorimetric

Compatible Sample Types: