Item no. |
PEL-MAPK-SK-2 |
Manufacturer |
Raybiotech
|
Amount |
2 Plate Kit |
Quantity options |
1 Plate Kit
2 Plate Kit
5 Plate Kit
|
Category |
|
Type |
Elisa-Kit |
Specific against |
other |
Host |
Hamster - Armenian |
ECLASS 10.1 |
32160605 |
ECLASS 11.0 |
32160605 |
UNSPSC |
41116126 |
Available |
|
Specificity |
The antibody pair provided in this kit recognizes ERK1/2 (T202/Y204), JNK (T183/Y185), P38&alpha, (T180/Y182), Total ERK1/2, JNK and P38&alpha, in Human, Mouse and Rat Cell Lysates. |
Species Detected |
Human Mouse Rat |
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 |
ANDERK1ERK2MAPK1MAPK3PRKM1PRKM2PRKM3 |
Kit Components |
1 Microplate ( Item A) Wash Buffer Concentrate (20x) ( Item B) Assay Diluent ( Item E) Detection Antibody Erk1/2( T202/ Y204) ( Item C-1) Detection Antibody Erk1/2 ( Item C-2) Detection Antibody J N K ( T183/ Y185) ( Item C-3) Detection Antibody J N K ( Item C-4) Detection Antibody P38α ( T180/ Y182) ( Item C-5) Detection Antibody P38α ( Item C-6) H R P-conjugated Anti-rabbit Ig G ( Item D-1) H R P-conjugated Streptavidin ( Item D-2) T M B One- Step Substrate Reagent ( Item H) Stop Solution ( Item I) Cell Lysate Buffer ( Item J) Positive Control A431 S002-1 ( Item K) |
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. |
Storage/Stability |
Upon receipt, the kit should be stored at â20°C. Please use within 6 months from the date of shipment. After initial use, Wash Buffer Concentrate (Item B), Assay Diluent (Item E), TMB One-Step Substrate Reagent (Item H), Stop Solution (Item I) and Cell Lysate Buffer (Item J) should be stored at 4°C to avoid repeated freeze-thaw cycles. Return unused wells to the pouch containing desiccant pack, reseal along entire edge, and store at â20°C. Item D, store at 2-8°C for up to one month (store at -20°C for up to 6 months, avoid repeated freeze-thaw cycles). Reconstituted Positive Control (Item K) should be stored at -70°C. |
Protein Name & Synonyms |
Mitogen-activated protein kinase 3 (MAP kinase 3) (MAPK 3) (EC 2.7.11.24) (ERT2) (Extracellular signal-regulated kinase 1) (ERK-1) (Insulin-stimulated MAP2 kinase) (MAP kinase isoform p44) (p44-MAPK) (Microtubule-associated protein 2 kinase) (p44-ERK1) Mitogen-activated protein kinase 1 (MAP kinase 1) (MAPK 1) (EC 2.7.11.24) (ERT1) (Extracellular signal-regulated kinase 2) (ERK-2) (MAP kinase isoform p42) (p42-MAPK) (Mitogen-activated protein kinase 2) (MAP kinase 2) (MAPK 2) |
Pathway |
MAPK Signaling |
Introduction |
RayBio® MAPK Pathway ELISA Sample 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 ERK1/2, JNK and P38α 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-ERK1/2, total ERK1/2, phospho-JNK, total JNK, phospho- P38α and total P38α. For each target, a capture antibody has been coated onto microwells. Samples are pipetted into the wells and target protein present in a sample is bound to the wells by the immobilized antibody. The wells are washed and a detection antibody is used to detect the captured target protein. After washing away unbound antibody, an HRP-conjugated secondary antibody 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 target protein bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm. |
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.