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Human/Mouse/Rat Phospho-NF-kB P65 (Ser536) and Total NF-kB P65 Cell-Based ELISA Kit

Phospho-NF-kB P65 (Ser536) and Total NF-kB P65 Cell-Based ELISA Kit. This assay semi-quantitatively measures NF-kB P65 phosphorylated at Serine-536 as well as total NF-kB P65 in adherent cell lines.

Product Description

Specifications

Size	1 Plate Kit, 2 Plate Kit, 5 Plate Kit
Estimated Lead Time	1-2 business days
Species	Human, Mouse, Rat
Specificity	This ELISA kit recognizes Human
Compatible Sample Types	Adherent Cell Culture
Accession Number	Q04206\|Q04207\|Q7TQN4
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)
Quantitative/Semi-Quantitative	Semi-Quantitative
Solid Support	96-well Microplate
Design Principle	Cell-based
Method Of Detection	Colorimetric
Research Area	Post-Translational Modifications, Phosphorylation, IGF Signaling, JAK/STAT Signaling, MAPK Signaling, mTOR Signaling, PI3K-AKT Signaling, NFkB Signaling, PKC Signaling
Shipping Type	Blue ice
Storage	-20°C

Product Features

Rapidly measure phosphorylated protein in adherent cell lines
Simultaneously measure Phosphorylated protein and pan protein in one experiment (for normalization purpose)
No sample lysis is needed
Compatible with a standard ELISA plate reader

Application Notes

Kit Components

Uncoated 96-well Strip Microplate
Wash Buffers
Fixing Solution
Quenching Buffer
Blocking Buffer
Anti-phospho antibody
Anti-pan antibody
HRP-Conjugated Secondary Antibody
TMB One-Step Substrate
Stop Solution

Other Materials Required

Distilled or deionized water
100 ml and 1 liter graduated cylinders
Tubes to prepare sample dilutions
Protease and Phosphatase inhibitors
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

Protocol Outline

Seed 10,000-30,000 cells into each well and incubate overnight.
Apply various treatment, inhibitors or activators according to manufacturer's instructions.
Add 100 µl of Fixing Solution into each well and incubate for 20 min at RT with shaking.
Add 200 µl of prepared 1X Quenching Buffer and incubate 20 min at RT.
Add 200 µl of Blocking Solution and incubate for 1 h at 37^°C.
Add 50 µl of 1X anti-phospho-protein specific antibody or anti-pan-protein specific antibody to each well and incubate for 2 h at RT.
Add 50 µl of prepared 1X HRP-Anti-Rabbit or Mouse IgG and incubate for 1 h at RT.
Add 100 µl of TMB One-Step Substrate Reagent to each well.
Incubate 30 min at RT.
Add 50 µl of Stop Solution to each well.
Read at 450 nm immediately.

Storage/Stability

Upon receipt, the kit should be stored at â20^°C. Please use within 6 months from the date of shipment.

Have this product in hand?

Get your lot-specific datasheet at Ray's Docs. The datasheet linked below is our current version and may not match your lot.

Datasheet (CBEL-NFKBP65-S536.pdf)

SDS (CBEL-NFKBP65-S536.pdf)

Product Features

Rapidly measure phosphorylated protein in adherent cell lines
Simultaneously measure Phosphorylated protein and pan protein in one experiment (for normalization purpose)
No sample lysis is needed
Compatible with a standard ELISA plate reader

Application Notes

Kit Components

Uncoated 96-well Strip Microplate
Wash Buffers
Fixing Solution
Quenching Buffer
Blocking Buffer
Anti-phospho antibody
Anti-pan antibody
HRP-Conjugated Secondary Antibody
TMB One-Step Substrate
Stop Solution

Other Materials Required

Distilled or deionized water
100 ml and 1 liter graduated cylinders
Tubes to prepare sample dilutions
Protease and Phosphatase inhibitors
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

Protocol Outline

Seed 10,000-30,000 cells into each well and incubate overnight.
Apply various treatment, inhibitors or activators according to manufacturer's instructions.
Add 100 µl of Fixing Solution into each well and incubate for 20 min at RT with shaking.
Add 200 µl of prepared 1X Quenching Buffer and incubate 20 min at RT.
Add 200 µl of Blocking Solution and incubate for 1 h at 37^°C.
Add 50 µl of 1X anti-phospho-protein specific antibody or anti-pan-protein specific antibody to each well and incubate for 2 h at RT.
Add 50 µl of prepared 1X HRP-Anti-Rabbit or Mouse IgG and incubate for 1 h at RT.
Add 100 µl of TMB One-Step Substrate Reagent to each well.
Incubate 30 min at RT.
Add 50 µl of Stop Solution to each well.
Read at 450 nm immediately.

Storage/Stability

Upon receipt, the kit should be stored at â20^°C. Please use within 6 months from the date of shipment.

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25% off all ELISA kits

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Human/Mouse/Rat Phospho-NF-kB P65 (Ser536) and Total NF-kB P65 Cell-Based ELISA Kit