Cell-Based Human/Mouse/Rat Stat 3 (Tyr705) Phosphorylation ELISA Kit 2 x 96-Well Kit
Antigen Information
Accession Number:
- P40763
Gene ID:
- 6774
Gene Symbol:
- APRF
- STAT3
Protein Name & Synonyms
Signal transducer and activator of transcription 3 (Acute-phase response factor)
Assay Format
Pathway
- JAK/STAT Signaling
Specificity
The antibodies provided in this kit recognizes human, mouse and rat Stat 3 phosphorylated at site Tyrosine-705 and total Stat 3 for comparison.
Number of Targets Detected
1
Species Detected:
- Human
- Mouse
- Rat
Compatible Sample Types:
- Adherent Cell Culture
Design Principle
- Cell-based
Method of Detection
Colorimetric
Quantitative/Semi-Quantitative:
Semi-Quantitative
Solid Support:
96-well Microplate
Product Specifications
Size
1, 2, or 5 x 96-Well Microplate Kit
Product Features
- Site and signal
Pathway
-specific
- In vitro detection of adherent cell culture
- No sample lysis is needed
- Compatible with a standard ELISA plate reader
- Faster results than with ELISA
- Adaptable for high-throughput screening and drug discovery
Application Notes
Kit Components
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 manufacture’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
The entire kit may be stored at -20°C for up to 6 months from the date of shipment. Avoid repeated freeze-thaw cycles.
Usage
This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.
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