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| Description | |
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| Catalogue Number | 15-107 |
| Brand Family | Upstate |
| Trade Name |
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| Description | MAPK Pathway 1 Explorer Antibody MiniPack |
| Alternate Names |
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| Background Information | Pathway Explorer Antibody MiniPack: Each Pathway Explorer Antibody Minipack contains three related antibodies as part of a signaling cascade or a combination of total and phosphorylated forms of key signaling targets. Each of the three antibodies are 30% the original pack size. Full size versions of each of the Pathway Explorer antibodies are available for sale individually under the same catalog number with the removal of “SP” off of each one (e.g. 05-591SP can be ordered as 05-591). Erk1/2: Erk (Extracellular signal-Related Kinase) is a family of two, highly homologous proteins denoted as Erk1 (p44, MAPK3) and Erk2 (p42, MAPK1) that both function in the same pathway. The two proteins are often referred to collectively as Erk1/2 or p44/p42 MAP kinase. The Erk pathway is considered the classical, canonical MAPK (Mitogen-Activated Protein Kinase) signaling pathway. It is an evolutionarily conserved pathway that controls and is a critical regulator of the growth and survival through the promotion of cell proliferation and the prevention of apoptosis. Erk is involved in the control of many fundamental cellular processes including cell proliferation, survival, differentiation, apoptosis, motility and metabolism. Erk is activated by growth factor stimulation of receptor tyrosine kinases (RTKs), GPCR, and/or integrin stimulation. This activates the Ras-Raf-MEK-Erk pathway that results in the phosphorylation/activation of Erk1/2 (p44/p42) on the TxY motif (Thr202/Tyr204 and Thr185/Tyr187 for Erk1 & Erk2, respectively). Raf: The Raf proteins (A-Raf, B-Raf, Raf-1/c-Raf) are Ser/Thr kinases with homology to the PKC family, containing an N-terminal regulatory domain and a C-terminal catalytic domain. Raf (MAP Kinase Kinase Kinase) is the top kinase in the canonical MAPK pathway. Members of the Raf family bind to activated Ras. Ras, in its GTP-bound active state, activates the kinase activity of Raf. This results in Raf translocation to the plasma membrane and activation. Once activated, Raf then binds to and activates MEK (MAP Kinase Kinase) by phosphorylating it on the two residue motif. It is thought that B-Raf might be the predominant activator of MEK and that Raf-1 has a role in protection against apoptosis; a process that does not require either its kinase activity or its activation of MEK. The regulation of a large number of cellular processes are dependent upon the activation state of ERK, so the controlling of this pathway could have profound effects on various diseases. Activation of Raf-1 involves phosphorylation of Ser338/339 and Tyr340/341. Activating mutations of B-Raf that disrupt its auto-inhibition loop have been implicated in a number of cancers, including melanoma and colon cancer. MEK1/MEK2: MAP Kinase/Erk Kinase (MEK), alternatively known as MKK, is a true dual-specificity kinase, in that it phosphorylates the MAP kinases (Erk1/2; p44/p42)) on both the Thr and Tyr of the activation motif TEY. In vitro, the tyrosine phosphorylation is favored, whereas in vivo both phosphorylation events appear to occur simultaneously. This suggests that an additional factor is present in cells to facilitate the reaction. MEK1 and MEK2 are activated by phosphorylation of two serine residues (Ser218/222 in MEK1 and Ser222/226 in MEK2), which are substrates for the Raf family of kinases. Mutation of the phosphorylation sites from Ser to Asp creates a protein with constitutive kinase activity, which when expressed in cells is able to cause transformation. * See full size versions for corresponding references. |
| Product Information | |
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| Components |
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| Format | Purified |
| Control |
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| Presentation | 3 individual tubes containing either Anti-phospho-Raf-1 (Ser338); or Anti-phospho-MEK1 (Ser218/222)/ MEK2 (Ser222/226); Anti-phospho-Erk 1/2 (Thr202/Tyr204, Thr185/Tyr187), recombinant clone AW39R |
| Properties | Each vial is 30% the size of the parent catalog number |
| Quality Level | MQ100 |
| Applications | |
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| Application | This Antibody pack contains Anti-phospho-Raf-1 Antibody(Ser338), Anti-phospho-MEK1 Antibody(Ser218/222)/ MEK2 (Ser222/226), Anti-phospho-Erk 1/2 Antibody (Thr202/Tyr204, Thr185/Tyr187), validated for use in IF, Flow, WB, Multiplexing |
| Key Applications |
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| Application Notes | 05-797RSP: Immunofluorescence Analysis: A431 cells were fixed, permabilized and dual stained with anti-phospho-Erk. Flow Cytometry Analysis:Jurkat cells were fixed, permeablized, and stained with anti-phospho-Erk 1/2 (Thr202/Tyr204, Thr185/Tyr187). |
| Biological Information | |
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| Immunogen | 05-797RSP: Phosphorylated peptide encompassing the TxY motif in the activation loop of Erk1 and Erk2.; 05-747SP: KLH-conjugated, synthetic peptide containing …DpSMANpSF… in which pS corresponds to phosphorylated amino acids 218 and 222 of human MEK1. The immunizing sequence is highly conserved.;05-538SP:KLH-conjugated, synthetic peptide encompassing and including phosphorylated Ser338 of human Raf-1. The immunizing sequence is identical in rat and chicken and has 12/13 identical amino acids in Xenopus. |
| Epitope | Please see relevant data sheet. |
| Clone | 05-797RSP: AW39R |
| Concentration | 05-538SP: 1 mg/mL |
| Specificity | 05-797RSP: Recognizes Erk 1 & 2 only when dually phosphorylated on its TxY activation motif.; 05-747SP: Phosphorylated MEK1/MEK2; 05-538SP: Recognizes Raf-1 phosphorylated at Serine 338 |
| Species Reactivity |
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| Species Reactivity Note | 05-538SP: Mouse and human. Predicted to cross react with rat, chicken and Xenopus based on sequence homology. |
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| Modifications |
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| UniProt Number |
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| UniProt Summary | Raf-1: FUNCTION: Involved in the transduction of mitogenic signals from the cell membrane to the nucleus. Part of the Ras-dependent signaling pathway from receptors to the nucleus. Protects cells from apoptosis mediated by STK3. CATALYTIC ACTIVITY: ATP + a protein = ADP + a phosphoprotein. COFACTOR: Binds 2 zinc ions per subunit. SUBUNIT STRUCTURE: Interacts with Ras proteins. The interaction with Ras is antagonized by RIN1. Weakly interacts with RIT1 By similarity. Interacts with STK3, which inhibits its pro-apoptotic activity. TISSUE SPECIFICITY: In skeletal muscle, isoform 1 is more abundant than isoform 2. POST-TRANSLATIONAL MODIFICATION: Phosphorylated upon DNA damage, probably by ATM or ATR. Phosphorylation at Thr-269 increases its kinase activity. INVOLVEMENT IN DISEASE: Defects in RAF1 are the cause of Noonan syndrome type 5 (NS5) [MIM:611553]. Noonan syndrome (NS) is a disorder characterized by dysmorphic facial features, short stature, hypertelorism, cardiac anomalies, deafness, motor delay, and a bleeding diathesis. It is a genetically heterogeneous and relatively common syndrome, with an estimated incidence of 1 in 1000-2500 live births. Ref.24 Ref.25 Defects in RAF1 are the cause of LEOPARD syndrome type 2 (LEOPARD syndrome-2) [MIM:611554]. LEOPARD syndrome is an autosomal dominant disorder allelic with Noonan syndrome. The acronym LEOPARD stands for lentigines, electrocardiographic conduction abnormalities, ocular hypertelorism, pulmonic stenosis, abnormalities of genitalia, retardation of growth, and deafness. MEK1: FUNCTION: Catalyzes the concomitant phosphorylation of a threonine and a tyrosine residue in a Thr-Glu-Tyr sequence located in MAP kinases. Activates ERK1 and ERK2 MAP kinases. CATALYTIC ACTIVITY: ATP + a protein = ADP + a phosphoprotein. ENZYME REGULATION: Activated by phosphorylation. SUBUNIT: Interacts with MORG1 (By similarity). Interacts with Yersinia yopJ. PTM: Phosphorylation on Ser/Thr by MAP kinase kinase kinases (RAF or MEKK1) regulates positively the kinase activity. PTM: Acetylation by Yersinia yopJ prevents phosphorylation and activation, thus blocking the MAPK signaling pathway. DISEASE: Defects in MAP2K1 are a cause of cardiofaciocutaneous syndrome (CFC syndrome) [MIM:115150]; also known as cardio-facio-cutaneous syndrome. CFC syndrome is characterized by a distinctive facial appearance, heart defects and mental retardation. Heart defects include pulmonic stenosis, atrial septal defects and hypertrophic cardiomyopathy. Some affected individuals present with ectodermal abnormalities such as sparse, friable hair, hyperkeratotic skin lesions and a generalized ichthyosis-like condition. Typical facial features are similar to Noonan syndrome. They include high forehead with bitemporal constriction, hypoplastic supraorbital ridges, downslanting palpebral fissures, a depressed nasal bridge, and posteriorly angulated ears with prominent helices. The inheritance of CFC syndrome is autosomal dominant. SIMILARITY: Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. MAP kinase kinase subfamily. SIMILARITY: Contains 1 protein kinase domain. Erk2: FUNCTION: Catalyzes the concomitant phosphorylation of a threonine and a tyrosine residue in a Thr-Glu-Tyr sequence located in MAP kinases. Activates ERK1 and ERK2 MAP kinases. CATALYTIC ACTIVITY: ATP + a protein = ADP + a phosphoprotein. ENZYME REGULATION: Activated by phosphorylation on tyrosine and threonine in response to insulin and NGF. Both phosphorylations are required for activity SUBUNIT STRUCTURE: Interacts with MORG1 By similarity. Binds to HIV-1 Nef through its SH3 domain. This interaction inhibits its tyrosine-kinase activity. Interacts with its substrates HSF4 and ARHGEF2. Interacts with NISCH. DOMAIN: The TXY motif contains the threonine and tyrosine residues whose phosphorylation activates the MAP kinases. PTM: Dually phosphorylated on Thr-185 and Tyr-187, which activates the enzyme. |
| Molecular Weight | 05-797RSP: 42 and 44 kDa; 05-747SP: 45 kDa; 05-538SP: 74 kDa |
| Product Usage Statements | |
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| Usage Statement |
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| Storage and Shipping Information | |
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| Storage Conditions | Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. |
| Packaging Information | |
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| Material Size | 3 vials/Pk |