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Phospho-SMAD2 (SER245/250/255) and Total SMAD2 Kit

Phospho-SMAD2 (SER245/250/255) and Total SMAD2 Kit

Item no.	PEL-SMAD2-S245-T-2
Manufacturer	Raybiotech
Amount	2 Plate Kit
Quantity options	1 Plate Kit 2 Plate Kit 5 Plate Kit
Category	Neuroscience Neural Lineage Markers ELISA ELISA & Immunoassays ELISA Kits Immunoassay Neurodegeneration Markers
Type	Elisa-Kit
Specific against	Human (Homo sapiens), Mouse (Murine, Mus musculus), Rat (Rattus norvegicus)
Host	Hamster - Armenian
ECLASS 10.1	32160605
ECLASS 11.0	32160605
UNSPSC	41116126
Shipping Condition	Cool pack
Available
Specificity	The antibody pair provided in this kit recognizes human, mouse, rat SMAD2 phosphorylated at site Serine-245/250/255 and Total SMAD2.
Manufacturer - Category	Assay Kits\|Phosphorylation Assays\|ELISA Kits\|Phosphorylation ELISA
Shipping Temperature	Blue ice
Storage Conditions	-20°C
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	MADH2MADR2SMAD2
Kit Components	Pre-Coated 96-well Strip Microplate Wash Buffer Anti-Phospho Antibody Anti-Pan Antibody HRP-Conjugated Secondary Antibody Streptavidin-Conjugated HRP Assay Diluent TMB One-Step Substrate Stop Solution Lysis Buffer Positive Control Sample
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	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 RT or O/N at 4°C. Add 100 µl of prepared primary antibody to each well. Incubate 1 h at RT. Add 100 µl of prepared 1 X HRP-Streptavidin to each well. Incubate 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.
Protein Name & Synonyms	Mothers against decapentaplegic homolog 2 (MAD homolog 2) (Mothers against DPP homolog 2) (JV18-1) (Mad-related protein 2) (hMAD-2) (SMAD family member 2) (SMAD 2) (Smad2) (hSMAD2)
UNSPSC Code	41116158
Manufacturer - Specificity	The antibody pair provided in this kit recognizes human, mouse, rat SMAD2 phosphorylated at site Serine-245/250/255 and Total SMAD2.
Pathway	AngiogenesisStem CellTGF-beta Signaling
Introduction	RayBio® Phospho-SMAD2 (Ser245/250/255) and Total SMAD2 ELISA kit is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in human, mouse and rat cell lysates. By determining phosphorylated SMAD2 protein 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 Blotting analysis. This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human, mouse and rat phospho-SMAD2 and total SMAD2. An anti-pan SMAD2 antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and SMAD2 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and rabbit anti-SMAD2 (Ser245/250/255) antibody is used to detect phosphorylated SMAD2 or mouse anti-SMAD2 antibody is used to detect pan SMAD2. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG or HRP-conjugated anti-mouse IgG 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 SMAD2 (Ser245/250/255) or pan SMAD2 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
Positive Control	Jurkat cells were treated with PMA. Solubilize cells at 4 x 107 cells/ml in Cell Lysate Buffer. Serial dilutions of lysates were analyzed in this ELISA. Please see step 3 of Part VI Reagent Preparation for detail.
Stimulation	Jurkat cells were treated or untreated with PMA. Cell lysates were analyzed using this phosphoELISA and Western Blot.

Note: The presented information and documents (Manual, Product Datasheet, Safety Datasheet and Certificate of Analysis) correspond to our latest update and should serve for orientational purpose only. We do not guarantee the topicality. We would kindly ask you to make a request for specific requirements, if necessary.

All products are intended for research use only (RUO). Not for human, veterinary or therapeutic use.

PEL-SMAD2-S245-T-2.pdf

PEL-SMAD2-S245-T-2-datasheet.pdf

PEL-SMAD2-S245-T-2-safety-datasheet.pdf