Western Blot (WB)
(Western Immunoblottlng solutions:Wash buffer: 1x Tris Buffered Saline (TBS): 0.1% Triton X-100Blocking buffer: 1xTBS: 0.1% Triton X-100: 5% BSA (used with the primary antibody))
Rabbit PHOSPHO-Erk 1,2 Monoclonal Antibody | anti-PHOSPHO-Erk 1,2 antibody
Anti - PHOSPHO-Erk 1,2
Tested: Rat
Tested: Rat
ELISA: 1:20000 - 1:100000
Immunoprecipitation to be tested by user.
Western Blot (WB)
(Western Immunoblottlng solutions:Wash buffer: 1x Tris Buffered Saline (TBS): 0.1% Triton X-100Blocking buffer: 1xTBS: 0.1% Triton X-100: 5% BSA (used with the primary antibody))
Western Blot (WB)
(Western Immunoblottlng solutions:Wash buffer: 1x Tris Buffered Saline (TBS): 0.1% Triton X-100Blocking buffer: 1xTBS: 0.1% Triton X-100: 5% BSA (used with the primary antibody))
Immunocytochemistry (ICC)
(1. Coat coverslips with 1 % gelatin-coating solution for 2 hours at room temperature (RT): rinse with distilled water, and let to dry overnight. Before plating the cells, wash the coated coverslips briefly with PBS.2. Fix the cells with 4% paraformaldehyde solution (in PBS, pH 7.2), for 15 min at RT.3. Wash 2 x 3 min with PBS.4. Permeabilize the cells with 0.1% Triton X-100 solution (in PBS. pH 7.2) for 5 min on ice.5. Wash 2 x 3 min with PBS.6. Incubate the cells in blocking buffer (0.3M glycine in PBS, 2% BSA) for 30 min at RT.7. Incubate the cells with primary antibody: anti-phospho Erk 1,2 clonal antibody at the dilution of 1:100 - 1 :400 in antibody dilution buffer (PBS, 1 % BSA) for 1 hour at RT in humid chamber.8. Wash 2 x 3 min with PBS.9. Apply the secondary antibody (in this case, the goat anti-rabbit IgG-FITC from Jackson Immunoresearch, cat. # 111-095-003, was used at 1 :300 in antibody dilution buffer, and cells were incubated for 1 hour at RT in dark).10. Wash 3 x 3 min with PBS.11. Rinse once with distilled water. 12. Mount the slide for observation, with a drop of anti-fade mounting medium.)
Immunocytochemistry (ICC)
(1. Coat coverslips with 1 % gelatin-coating solution for 2 hours at room temperature (RT): rinse with distilled water, and let to dry overnight. Before plating the cells, wash the coated coverslips briefly with PBS.2. Fix the cells with 4% paraformaldehyde solution (in PBS, pH 7.2), for 15 min at RT.3. Wash 2 x 3 min with PBS.4. Permeabilize the cells with 0.1% Triton X-100 solution (in PBS. pH 7.2) for 5 min on ice.5. Wash 2 x 3 min with PBS.6. Incubate the cells in blocking buffer (0.3M glycine in PBS, 2% BSA) for 30 min at RT.7. Incubate the cells with primary antibody: anti-phospho Erk 1,2 clonal antibody at the dilution of 1:100 - 1 :400 in antibody dilution buffer (PBS, 1 % BSA) for 1 hour at RT in humid chamber.8. Wash 2 x 3 min with PBS.9. Apply the secondary antibody (in this case, the goat anti-rabbit IgG-FITC from Jackson Immunoresearch, cat. # 111-095-003, was used at 1 :300 in antibody dilution buffer, and cells were incubated for 1 hour at RT in dark).10. Wash 3 x 3 min with PBS.11. Rinse once with distilled water. 12. Mount the slide for observation, with a drop of anti-fade mounting medium.)
Basic Information: Major clone of IgGs representing both, Erk1 and Erk2, obtained from immunoglobulins corresponding to pTE-pY dual phosphorylation sequence
NCBI and Uniprot Product Information
NCBI Description
This gene encodes a member of the MAP kinase family. MAP kinases, also known as extracellular signal-regulated kinases (ERKs), act as an integration point for multiple biochemical signals, and are involved in a wide variety of cellular processes such as proliferation, differentiation, transcription regulation and development. The activation of this kinase requires its phosphorylation by upstream kinases. Upon activation, this kinase translocates to the nucleus of the stimulated cells, where it phosphorylates nuclear targets. One study also suggests that this protein acts as a transcriptional repressor independent of its kinase activity. The encoded protein has been identified as a moonlighting protein based on its ability to perform mechanistically distinct functions. Two alternatively spliced transcript variants encoding the same protein, but differing in the UTRs, have been reported for this gene. [provided by RefSeq, Jan 2014]
Uniprot Description
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 plays also 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, DCC, 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. Mediates phosphorylation of TPR in respons to EGF stimulation. May play a role in the spindle assembly checkpoint. Phosphorylates PML and promotes its interaction with PIN1, leading to PML degradation. Ref.7 Ref.8 Ref.10 Ref.11 Ref.12 Ref.13 Ref.15 Ref.16 Ref.17 Ref.18 Ref.19 Ref.21 Ref.22 Ref.23 Ref.26 Ref.27 Ref.28 Ref.29 Ref.30 Ref.31 Ref.34 Ref.40 Ref.43 Ref.44 Ref.55Acts as a transcriptional repressor. Binds to a [GC]AAA[GC] consensus sequence. Repress the expression of interferon gamma-induced genes. Seems to bind to the promoter of CCL5, DMP1, IFIH1, IFITM1, IRF7, IRF9, LAMP3, OAS1, OAS2, OAS3 and STAT1. Transcriptional activity is independent of kinase activity. Ref.7 Ref.8 Ref.10 Ref.11 Ref.12 Ref.13 Ref.15 Ref.16 Ref.17 Ref.18 Ref.19 Ref.21 Ref.22 Ref.23 Ref.26 Ref.27 Ref.28 Ref.29 Ref.30 Ref.31 Ref.34 Ref.40 Ref.43 Ref.44 Ref.55
Catalytic activity: ATP + a protein = ADP + a phosphoprotein.
Cofactor: Magnesium
By similarity.
Enzyme regulation: Phosphorylated by MAP2K1/MEK1 and MAP2K2/MEK2 on Thr-185 and Tyr-187 in response to external stimuli like insulin or NGF. Both phosphorylations are required for activity. This phosphorylation causes dramatic conformational changes, which enable full activation and interaction of MAPK1/ERK2 with its substrates. Phosphorylation on Ser-29 by SGK1 results in its activation by enhancing its interaction with MAP2K1/MEK1 and MAP2K2/MEK2. Dephosphorylated and inactivated by DUSP3, DUSP6 and DUSP9. Inactivated by pyrimidylpyrrole inhibitors. Ref.19 Ref.48
Subunit structure: Binds both upstream activators and downstream substrates in multimolecular complexes. Binds to HIV-1 Nef through its SH3 domain. This interaction inhibits its tyrosine-kinase activity. Interacts with ADAM15, ARHGEF2, ARRB2, DAPK1 (via death domain), HSF4, IER3, IPO7, DUSP6, NISCH, SGK1, and isoform 1 of NEK2. Interacts (phosphorylated form) with CAV2 ('Tyr-19'-phosphorylated form); the interaction, promoted by insulin, leads to nuclear location and MAPK1 activation. Interacts with MORG1, PEA15 and MKNK2
By similarity. MKNK2 isoform 1 binding prevents from dephosphorylation and inactivation
By similarity. Interacts with DCC
By similarity. The phosphorylated form interacts with PML (isoform PML-4) Ref.9 Ref.13 Ref.19 Ref.20 Ref.24 Ref.29 Ref.33 Ref.34 Ref.36 Ref.37 Ref.39 Ref.40 Ref.46 Ref.48 Ref.55
Subcellular location: Cytoplasm › cytoskeleton › spindle
By similarity. Nucleus. Cytoplasm › cytoskeleton › microtubule organizing center › centrosome. Cytoplasm. Note: Associated with the spindle during prometaphase and metaphase
By similarity. PEA15-binding and phosphorylated DAPK1 promote its cytoplasmic retention. Phosphorylation at Ser- 246 and Ser-248 as well as autophosphorylation at Thr-190 promote nuclear localization. Ref.24 Ref.29 Ref.39 Ref.48
Domain: The TXY motif contains the threonine and tyrosine residues whose phosphorylation activates the MAP kinases.
Post-translational modification: Phosphorylated upon KIT and FLT3 signaling
By similarity. Dually phosphorylated on Thr-185 and Tyr-187, which activates the enzyme. Undergoes regulatory phosphorylation on additional residues such as Ser-246 and Ser-248 in the kinase insert domain (KID) These phosphorylations, which are probably mediated by more than one kinase, are important for binding of MAPK1/ERK2 to importin-7 (IPO7) and its nuclear translocation. In addition, autophosphorylation of Thr-190 was shown to affect the subcellular localization of MAPK1/ERK2 as well. Ligand-activated ALK induces tyrosine phosphorylation. Dephosphorylated by PTPRJ at Tyr-187. Phosphorylation on Ser-29 by SGK1 results in its activation by enhancing its interaction with MAP2K1/MEK1 and MAP2K2/MEK2. DUSP3 and DUSP6 dephosphorylate specifically MAPK1/ERK2 and MAPK3/ERK1 whereas DUSP9 dephosphorylates a broader range of MAPKs. Ref.14 Ref.35 Ref.39 Ref.44 Ref.45 Ref.46 Ref.48 Ref.63ISGylated
By similarity.
Sequence similarities: Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. MAP kinase subfamily.Contains 1 protein kinase domain.
Sequence caution: The sequence CAA77753.1 differs from that shown. Reason: Erroneous initiation. Translation N-terminally extended.
Research Articles on PHOSPHO-Erk 1,2
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Product Notes
The PHOSPHO-Erk 1,2 mapk1 (Catalog #AAA684145) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti - PHOSPHO-Erk 1,2 reacts with Human, Mouse Tested: Rat and may cross-react with other species as described in the data sheet. AAA Biotech's PHOSPHO-Erk 1,2 can be used in a range of immunoassay formats including, but not limited to, ELISA (EIA), Western Blot (WB), Immunoprecipitation (IP), Immunocytochemistry (ICC). WB: 1:5000 ELISA: 1:20000 - 1:100000 Immunoprecipitation to be tested by user. Researchers should empirically determine the suitability of the PHOSPHO-Erk 1,2 mapk1 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "PHOSPHO-Erk 1,2, Monoclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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