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Custom Human Phospho-IkB-alpha (Ser32) and Total IKB-alpha Cell-Based ELISA Kit
RayBio® Human Phospho-IkB-alpha (Ser32) and Total IKB-alpha Cell-Based ELISA Kit. This assay semi-quantitatively measures IKB-alpha phosphorylated at Serine-32 as well as total IKB-alpha in adherent cell lines.
Product Description
Specifications
Size
1 Plate Kit, 2 Plate Kit, 5 Plate Kit
Species
Human
Accession Number
P25963
Gene Id
4792
Gene Symbols
NFKBIA|IKBA|MAD3|NFKBI
Protein Name / Synonyms
NF-kappa-B inhibitor alpha (I-kappa-B-alpha) (IkB-alpha) (IkappaBalpha) (Major histocompatibility complex enhancer-binding protein MAD3)
Quantitative/Semi-Quantitative
Semi-Quantitative
Specificity
This ELISA kit recognizes Human IKB-alpha phosphorylated at site Serine-32 as well as total IKB-alpha.
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, mTOR Signaling, NFkB Signaling, PKC Signaling
Estimated Lead Time
4-6 weeks
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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$5 Amazon gift card in every kit box purchased.
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.
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