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.
|Detection Range||78.13-5000 pg/mL|
|Sample Volume Required Per Well||100uL|
|Sample Type||Serum, 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.
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.
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|
|C V (%)||6.74||5.35||4.59||6.36||5.17||5.19|
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 (%)|
|EDTA plasma (n=5)||92-105||98|
|Cell culture media (n=5)||91-103||96|
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)|
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.
|Micro ELISA Plate(Dismountable)||8 wells X12 strips||-20'C, 6 months|
|Reference Standard||2 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 Diluent||1 vial, 20 mL||4'C, 6 months|
|Biotinylated Detection Ab Diluent||1 vial, 14 mL|
|HRP Conjugate Diluent||1 vial, 14 mL|
|Concentrated Wash Buffer (25X)||1 vial, 30 mL|
|Substrate Reagent||1 vial, 10 mL||4'C(shading light)|
|Stop Solution||1 vial, 10 mL||4'C|
|Plate Sealer||5 pieces|
|Product Description||1 copy|
|Certificate of Analysis||1 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 188.8.131.52; 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
|NCBI GenInfo Identifier:||52001483|
|NCBI Gene ID:||26417|
|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|