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	The antibodies provided in this kit recognizes human, mouse and rat Stat 3 phosphorylated at site Tyrosine-705 and total Stat 3 for comparison.
Compatible Sample Types	Adherent Cell Culture
Accession Number	P40763
Gene Id	6774
Gene Symbols	STAT3, APRF
Protein Name / Synonyms	Signal transducer and activator of transcription 3 (Acute-phase response factor)
Quantitative/Semi-Quantitative	Semi-Quantitative
Solid Support	96-well Microplate
Design Principle	Cell-based
Method Of Detection	Colorimetric
Research Area	Angiogenesis, Cancer, Post-Translational Modifications, Inflammation, Phosphorylation, JAK/STAT Signaling, Autophagy
Shipping Type	Blue ice
Storage	-20°C

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

uncoated 96-well Microplate

Wash Buffer A

Wash Buffer B

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 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.

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

uncoated 96-well Microplate

Wash Buffer A

Wash Buffer B

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 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.

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