ERK1 (phospho T202 + Y204) + ERK2 (phospho T185 + Y187) + Total ELISA Kit (RMEK-0151389)
Cat. No.: RMEK-0151389
Category: ELISA Kits
INQUIRY
1 x 96 tests
ERK1 (phospho T202 + Y204) + ERK2 (phospho T185 + Y187) + Total ELISA kit is designed for the semi-quantitative measurement of ERK1/2 (pT202/Y204) and Total ERK1/2 protein in Human and mouse cells.
The ELISA employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.
ASSAY SPECIFICITY
The ERK1/2 (pT202/Y204) assay detects endogenous levels of ERK1/2 (GenBank Accessions NP 002737. 2 [ERK1], NP 620407 [ERK2]) in cellular lysates, only when phosphorylated at Thr202/Tyr204.
The ERK1/2 Total assay detects endogenous levels of ERK1/2 (GenBank Accessions NP 002737. 2 [ERK1], NP 620407 [ERK2]) in cellular lysates, irrespective of phosphorylation status.
SPECIES REACTIVITY
This kit detects ERK1/2 (pT202/Y204) and ERK1/2 Total in Human and mouse cell culture extracts. Detection in rat samples is also expected. Other species should be tested on a case-by-case basis.
Serum and plasma samples have not been tested with this kit.
The ELISA employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.
ASSAY SPECIFICITY
The ERK1/2 (pT202/Y204) assay detects endogenous levels of ERK1/2 (GenBank Accessions NP 002737. 2 [ERK1], NP 620407 [ERK2]) in cellular lysates, only when phosphorylated at Thr202/Tyr204.
The ERK1/2 Total assay detects endogenous levels of ERK1/2 (GenBank Accessions NP 002737. 2 [ERK1], NP 620407 [ERK2]) in cellular lysates, irrespective of phosphorylation status.
SPECIES REACTIVITY
This kit detects ERK1/2 (pT202/Y204) and ERK1/2 Total in Human and mouse cell culture extracts. Detection in rat samples is also expected. Other species should be tested on a case-by-case basis.
Serum and plasma samples have not been tested with this kit.
Product Features
| Species Reactivity | Mouse, Human |
|---|---|
| Detection Method | Colorimetric |
| Assay Duration | One step assay |
| Assay Time | 1.5 h |
| Assay Type | Semi-quantitative |
| Precision | Intra-Assay-(pT202/Y204)-6-3.3%; Intra-Assay-(Total)-6-3%; Inter-Assay-(pT202/Y204)-3-1.3%; Inter-Assay-(Total)-3-4.9% |
| Sample Type | Cell Lysate, Tissue Homogenate |
| Key Features | One-wash 90 minute protocol; Sample type: Cell Lysate, Tissue Homogenate; Detection method: Colorimetric; Assay type: Semi-quantitative; Reacts with: Mouse, Human |
Target Information
| Target Symbol | ERK1 |
|---|---|
| UniProt ID | P27361 |
| Function | Serine/threonine kinase which acts as an essential component of the MAP kinase signal transduction pathway. MAPK1/ERK2 and MAPK3/ERK1 are the 2 MAPKs which play an important role in the MAPK/ERK cascade. They participate also in a signaling cascade initiated by activated KIT and KITLG/SCF. Depending on the cellular context, the MAPK/ERK cascade mediates diverse biological functions such as cell growth, adhesion, survival and differentiation through the regulation of transcription, translation, cytoskeletal rearrangements. The MAPK/ERK cascade also plays a role in initiation and regulation of meiosis, mitosis, and postmitotic functions in differentiated cells by phosphorylating a number of transcription factors. About 160 substrates have already been discovered for ERKs. Many of these substrates are localized in the nucleus, and seem to participate in the regulation of transcription upon stimulation. However, other substrates are found in the cytosol as well as in other cellular organelles, and those are responsible for processes such as translation, mitosis and apoptosis. Moreover, the MAPK/ERK cascade is also involved in the regulation of the endosomal dynamics, including lysosome processing and endosome cycling through the perinuclear recycling compartment (PNRC); as well as in the fragmentation of the Golgi apparatus during mitosis. The substrates include transcription factors (such as ATF2, BCL6, ELK1, ERF, FOS, HSF4 or SPZ1), cytoskeletal elements (such as CANX, CTTN, GJA1, MAP2, MAPT, PXN, SORBS3 or STMN1), regulators of apoptosis (such as BAD, BTG2, CASP9, DAPK1, IER3, MCL1 or PPARG), regulators of translation (such as EIF4EBP1) and a variety of other signaling-related molecules (like ARHGEF2, DEPTOR, FRS2 or GRB10). Protein kinases (such as RAF1, RPS6KA1/RSK1, RPS6KA3/RSK2, RPS6KA2/RSK3, RPS6KA6/RSK4, SYK, MKNK1/MNK1, MKNK2/MNK2, RPS6KA5/MSK1, RPS6KA4/MSK2, MAPKAPK3 or MAPKAPK5) and phosphatases (such as DUSP1, DUSP4, DUSP6 or DUSP16) are other substrates which enable the propagation the MAPK/ERK signal to additional cytosolic and nuclear targets, thereby extending the specificity of the cascade. |
| Cellular Localization | Cytoplasm. Nucleus. Membrane, caveola. Cell junction, focal adhesion. Autophosphorylation at Thr-207 promotes nuclear localization. PEA15-binding redirects the biological outcome of MAPK3 kinase-signaling by sequestering MAPK3 into the cytoplasm (By similarity). |
Storage & Shipping
| Storage | Store at 2-8°C |
|---|---|
| Shipping | Gel Packs |
| Stability | The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. |
For research use only. Not for clinical use.
