Mouse ERK1 (Extracellular Signal Regulated Kinase 1) ELISA Kit (MOES01005)



ELISA Kit Technical ManualMSDS

Mouse ERK1 (Extracellular Signal Regulated Kinase 1) ELISA Kit

This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Mouse ERK1 . Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Mouse ERK1 and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Mouse ERK1, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Mouse ERK1. The concentration of Mouse ERK1 in samples can be calculated by comparing the OD of the samples to the standard curve.

Assay typeSandwich
Assay time4.5h
Detection MethodColormetric
Detection Range78.13-5000 pg/mL
Sensitivity46.88 pg/mL
Sample Volume Required Per Well100uL
Sample TypeSerum, plasma and other biological fluids


This kit recognizes Mouse ERK1 in samples. No significant cross-reactivity or interference between Mouse ERK1 and analogues was observed.

Typical Data

As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

O.D Average Corrected


Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse ERK1 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse ERK1 were tested on 3 different plates, 20 replicates in each plate.

Intra-assay Precision Inter-assay Precision
Mean (pg/mL)263.22572.261803.23281.37609.581846.30
Standard deviation17.7430.6282.7717.9031.5295.82
C V (%)6.745.354.596.365.175.19


The recovery of Mouse ERK1 spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type Range (%) Average Recovery (%)
Serum (n=5)95-108102
EDTA plasma (n=5)92-10598
Cell culture media (n=5)91-10396


Samples were spiked with high concentrations of Mouse ERK1 and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
1:2Range (%)93-11084-9788-102
Average (%)1019094
1:4Range (%)91-10782-9287-99
Average (%)988793
1:8Range (%)92-10382-9685-96
Average (%)978991
1:16Range (%)90-10281-9485-95
Average (%)968889

Kit Components & Storage

An unopened kit can be stored at 4'C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

Item Specifications Storage
Micro ELISA Plate(Dismountable)8 wells X12 strips-20'C, 6 months
Reference Standard2 vials
Concentrated Biotinylated Detection Ab (100X)1 vial, 120 uL
Concentrated HRP Conjugate (100X)1 vial, 120 uL-20'C(shading light), 6 months
Reference Standard & Sample Diluent1 vial, 20 mL4'C, 6 months
Biotinylated Detection Ab Diluent1 vial, 14 mL
HRP Conjugate Diluent1 vial, 14 mL
Concentrated Wash Buffer (25X)1 vial, 30 mL
Substrate Reagent1 vial, 10 mL4'C(shading light)
Stop Solution1 vial, 10 mL4'C
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 copy

Mouse ERK1 (Extracellular Signal Regulated Kinase 1) ELISA Kit (MOES01005) Assay procedure

    1. Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
    2. Aliquot 100µl of standard solutions into the standard wells.
    3. Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
    4. Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids) into test sample wells.
    5. Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
    6. Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix.Incubate for 1 hour at 37°C.
    7. Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
    8. Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
    9. Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
    10. Add 90µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37°C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
    11. Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
    12. Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.

Mouse ERK1 (Extracellular Signal Regulated Kinase 1) ELISA Kit (MOES01005) Protein Information

UniProt Protein Function:ERK1: a serine/threonine kinase of the GMGC group that plays a critical role in the regulation of cell growth and differentiation. ERK1 (MAPK3) and ERK2 (MAPK1) play central roles in MAPK cascades and are activated by a wide variety of extracellular signals including growth and neurotrophic factors, cytokines, hormones and neurotransmitters. Depending on the cellular context, MAPK cascades mediate diverse biological functions such as cell growth, adhesion, survival and differentiation through the regulation of transcription, translation, cytoskeletal rearrangements. MAPK cascades also plays a role in initiation and regulation of meiosis, mitosis, and postmitotic functions in differentiated cells by phosphorylating a number of transcription factors. Activation of MAP kinases occurs through phosphorylation of threonine and tyrosine residues at the sequence T*EY* by upstream MAP kinase kinases, MEK1 and -2. Phosphorylation of both the threonine and tyrosine are required for activity. This phosphorylation causes dramatic conformational changes, which enable full activation and interaction of MAPK1/ERK2 with its substrates.
UniProt Protein Details:

Protein type:Kinase, protein; EC; Protein kinase, CMGC; Protein kinase, Ser/Thr (non-receptor); CMGC group; MAPK family; MAPK/ERK subfamily; ERK subfamily

Cellular Component: caveola; cytoplasm; cytoskeleton; cytosol; early endosome; focal adhesion; Golgi apparatus; late endosome; microtubule cytoskeleton; mitochondrion; nuclear envelope; nucleoplasm; nucleus; protein complex; pseudopodium

Molecular Function:ATP binding; MAP kinase activity; phosphatase binding; phosphotyrosine binding; protein binding; protein kinase activity

Biological Process: Bergmann glial cell differentiation; BMP signaling pathway; cartilage development; DNA damage induced protein phosphorylation; lipopolysaccharide-mediated signaling pathway; MAPKKK cascade; neural crest cell development; nuclear translocation of MAPK; organ morphogenesis; outer ear morphogenesis; peptidyl-serine phosphorylation; phosphorylation; positive regulation of cyclase activity; positive regulation of histone acetylation; positive regulation of histone phosphorylation; positive regulation of protein amino acid phosphorylation; positive regulation of telomerase activity; positive regulation of telomere maintenance via telomerase; positive regulation of transcription from RNA polymerase II promoter; positive regulation of translation; protein amino acid phosphorylation; protein complex assembly; regulation of cytoskeleton organization and biogenesis; regulation of ossification; regulation of stress-activated MAPK cascade; regulation of transcription factor activity; response to DNA damage stimulus; response to exogenous dsRNA; response to lipopolysaccharide; response to toxin; sensory perception of pain; signal transduction; thymus development; thyroid gland development; transcription, DNA-dependent

UniProt Code:Q63844
NCBI GenInfo Identifier:52001483
NCBI Gene ID:26417
NCBI Accession:Q63844.5
UniProt Secondary Accession:Q63844,Q61531, Q8K0X5, Q91YW5,
UniProt Related Accession:Q63844
Molecular Weight:43,066 Da
NCBI Full Name:Mitogen-activated protein kinase 3
NCBI Synonym Full Names:mitogen-activated protein kinase 3
NCBI Official Symbol:Mapk3
NCBI Official Synonym Symbols:p44; Erk1; Ert2; Mnk1; Erk-1; Esrk1; Prkm3; Mtap2k; p44erk1; p44mapk
NCBI Protein Information:mitogen-activated protein kinase 3
UniProt Protein Name:Mitogen-activated protein kinase 3
UniProt Synonym Protein Names:ERT2; Extracellular signal-regulated kinase 1; ERK-1; Insulin-stimulated MAP2 kinase; MAP kinase isoform p44; p44-MAPK; MNK1; Microtubule-associated protein 2 kinase; p44-ERK1
UniProt Gene Name:Mapk3
UniProt Entry Name:MK03_MOUSE
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