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Human/Mouse Phospho-Stat 1 (Y701) Cell-Based ELISA

Human/Mouse Phospho-Stat 1 (Y701) Cell-Based ELISA

Manufacturer	Raybiotech
Category	ELISA ELISA & Immunoassays ELISA Kits Immunoassay
Type	Elisa-Kit
Specific against	Human, Mouse, Rat
Amount	5 Plate Kit
Host	Hamster - Armenian
Item no.	CBEL-STAT1-5
eClass 6.1	32160605
eClass 9.0	32160605
Available
Species Detected	Human Mouse
Compatible Sample Types	Adherent Cell Culture
Design Principle	Cell-based
Method of Detection	Colorimetric
Quantitative/Semi-Quantitative	Semi-Quantitative
Solid Support	96-well Microplate
Gene Symbols	STAT1
Kit Components	uncoated 96-well Microplate Wash Buffer A Wash Buffer B Fixing Solution Quenching Buffer Blocking Buffer Anti-phospho antibody Anti-pan antibody H R P- Conjugated Secondary Antibody T M B One- Step Substrate Stop Solution
Other Materials Required	Distilled or deionized water100 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 R T with shaking. Add 200 µl of prepared 1 X Quenching Buffer and incubate 20 min at R T. Add 200 µl of Blocking Solution and incubate for 1 h at 37 ° C. Add 50 µl of 1 X anti-phospho-protein specific antibody or anti-pan-protein specific antibody to each well and incubate for 2 h at R T. Add 50 µl of prepared 1 X H R P- Anti- Rabbit or Mouse Ig G and incubate for 1 h at R T. Add 100 µl of T M B One- Step Substrate Reagent to each well. Incubate 30 min at R T. Add 50 µl of Stop Solution to each well. Read at 450 nm immediately.
Protein Name & Synonyms	Signal transducer and activator of transcription 1-alpha/beta (Transcription factor ISGF-3 components p91/p84)
Pathway	JAK/STAT Signaling

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