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Human Phosphotyrosine BTK ELISA

Human Phosphotyrosine BTK ELISA

ArtNr	PEL-BTK-Y-5
Hersteller	Raybiotech
Menge	5 Plate Kit
Quantity options	1 x 96-Well Strip Microplate Kit 2 Plate Kit 5 Plate Kit
Kategorie	ELISA ELISA & Immunoassays ELISA Kits Immunoassay
Typ	Elisa-Kit
Specific against	Human
Host	Hamster - Armenian
ECLASS 10.1	32160605
ECLASS 11.0	32160605
UNSPSC	41116126
Similar products	Btk
Lieferbar
Specificity	The antibody pair provided in this kit recognizes Human Tyrosine-Phosphorylated-BTK.
Species Detected	Human
Compatible Sample Types	Cell Lysates Tissue Lysates
Design Principle	Sandwich-based
Method of Detection	Colorimetric
Quantitative/Semi-Quantitative	Semi-Quantitative
Solid Support	96-well Microplate
Gene Symbols	AGMX1ATKBPKBTK
Kit Components	Pre- Coated 96-well Strip Microplate Wash Buffer Biotinylated Anti- Phosphotyrosine Antibody Stop Solution Assay Diluent(s) Positive Control Sample Lysis Buffer Streptavidin- Conjugated H R P T M B One- Step Substrate
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	Prepare all reagents and samples as instructed in the manual. Add 100 µl of sample or positive control to each well. Incubate 2.5 h at R T or O/ N at 4 ° C. Add 100 µl of prepared primary antibody to each well. Incubate 1 h at R T. Add 100 µl of prepared 1 X H R P- Streptavidin to each well. Incubate 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	Tyrosine-protein kinase BTK (EC 2.7.10.2) (Agammaglobulinemia tyrosine kinase) (ATK) (B-cell progenitor kinase) (BPK) (Bruton tyrosine kinase)
Pathway	B Cell ReceptorTyrosine Kinase Family
Introduction	RayBio® Phospho-BTK ELISA kit is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in cell lysates. By determining phospho-BTK in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blot analysis.This sandwich-based ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human phospho-BTK. An anti-BTK antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and phosphorylated and unphosphorylated BTK present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-phosphotyrosine antibody is used to detect only tyrosine-phosphorylated protein. After washing away unbound antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of phosphor-BTK bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
Positive Control	THP1 cells were treated with Pervanadate at 37°C for 10 min. Solubilize cells at 4 x 107 cells/ml in lysis buffer. Serial dilutions of lysates were analyzed in this ELISA. Please see step 3 of Part VI Reagent Preparation for detail.
Pervanadate Stimulation of U937 Cell Line	U937 cells were treated or untreated with Pervanadate for 10 min at 37°C. Cell lysates were analyzed using this phosphoELISA

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