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Human, Mouse and 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
Species
Human, Mouse, Rat
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
Specificity
This ELISA kit recognizes Human
Compatible Sample Types
Adherent Cell Culture
Solid Support
96-well Microplate
Method Of Detection
Colorimetric
Design Principle
Cell-based
Research Area
Post-Translational Modifications, Phosphorylation, IGF Signaling, JAK/STAT Signaling, MAPK Signaling, mTOR Signaling, PI3K-AKT Signaling, NFkB Signaling, PKC Signaling
Estimated Lead Time
1-2 business days
Shipping Type
Blue ice
Storage
-20°C
Risk-Free Guarantee
We offer a 100% guarantee on all ELISA kits and membrane cytokine arrays.
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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 –20° C. Please use within 6 months from the date of shipment.
Easy ELISA
Easy to run on human proximal tubule cells. Data generated in 24 hrs. Instructions easy to follow. User friendly protocol.
Review by
MKasztan,
from
UAB,
on
May 21, 2024
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