Human/Mouse/Rat Phospho-Erk1 (T202/Y204) + Phospho-Erk2 (T185/Y187) and Total Erk1/2 PharmaGenie ELISA Kit (SBRS1823)

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Human/Mouse/Rat Phospho-Erk1 (T202/Y204) + Phospho-Erk2 (T185/Y187) and Total Erk1/2 PharmaGenie ELISA - Kit Data

Product SKU:SBRS1823
Application:The antibody pair provided in this kit recognizes human, mouse and rat Phospho-Erk1 (pThr202/pTyr204) + Erk2(Tyr185/187) and pan Erk1/2.
Uniprot:P27361 / P28482
Gene ID:5595 and 5594
Gene Names:AND / ERK1 / ERK2 / MAPK1 / MAPK3 / PRKM1 / PRKM2 / PRKM3
Pathway:MAPK Signaling
Synonyms:Mitogen-activated protein kinase 3 (MAP kinase 3) (MAPK 3) (EC (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 (ERT1) (Extracellular signal-regulated kinase 2) (ERK-2) (MAP kinase isoform p42) (p42-MAPK) (Mitogen-activated protein kinase 2) (MAP kinase 2) (MAPK 2)
Target Species:Human, Mouse, Rat
Compatible Sample Types:Cell Lysates, Tissue Lysates
Design Principle:Sandwich-based
Method of Detection:Colorimetric
Storage/Stability:Upon receipt, the kit should be stored at -20 °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), HRP-Streptavidin (Item G), 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 -20 °C. Reconstituted Positive Control (Item K) should be stored at -70 °C.

Kit Components:

1.Pre-Coated 96-well Strip Microplate
2.Wash Buffer
3.Anti-Phospho Antibody
4.Anti-Pan Antibody
5.HRP-Conjugated Secondary Antibody
6.Streptavidin-Conjugated HRP
7.Assay Diluent
8.TMB One-Step Substrate
9.Stop Solution
10.Lysis Buffer
11.Positive Control Sample

Other materials and equipment required:

The Assay Genie Human/Mouse/Rat Phospho-Erk1 (T202/Y204) + Phospho-Erk2 (T185/Y187) and Total Erk1/2 ELISA Kit will require other equipment and materials to carry out the assay. Please see list below for further details.

  • 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 ul 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

Kit Protocol:

1.Prepare all reagents and samples as instructed in the manual.
2.Add 100 ul of sample or positive control to each well.
3.Incubate 2.5 h at RT or O/N at 4 °C.
4.Add 100 ul of prepared primary antibody to each well.
5.Incubate 1 h at RT.
6.Add 100 ul of prepared 1X HRP-Streptavidin to each well.
7.Incubate 1 h at RT.
8.Add 100 ul of TMB One-Step Substrate Reagent to each well.
9.Incubate 30 min at RT.
10.Add 50 ul of Stop Solution to each well.
11.Read at 450 nm immediately.