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Extracts prepared from HEK293 cells transiently transfected with plasmids expressing ERK5 kinase domain (ERK5kin) and constitutively activated MEK5D-D were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4°C, then were incubated with the ERK5 [pTpY218/220] antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1), the non-phosphopeptide corresponding to the immunogen (2), a generic phosphothreonine-containing peptide (3), a generic phosphotyrosine-containing peptide (4), the phosphopeptide derived from the corresponding region of ERK1&2 (5), or, the phosphopeptide immunogen (6). After washing, membranes were incubated with goat F (ab"e;)2 anti-rabbit IgG alkaline phosphatase conjugate (Cat. no. ALI4405) and bands were detected using the Tropix WesternStar™ detection method. The data show that while there is some cross-reactivity with ERK1&2, only the phosphopeptide corresponding to ERK5 [pTpY218/220] completely blocks the antibody signal, demonstrating the specificity of the antibody. NOTE: The antibody signal appears at ~50 kDa as this is the molecular weight of the transiently transfected ERK5 kinase domain.
|Tested species reactivity||Human|
|Published species reactivity||Mouse, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from the region of human ERK5 that contains threonine 218 and tyrosine 220. The sequence is conserved in mouse.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 50% glycerol, 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||Assay Dependent|
|Immunofluorescence (IF)||Assay Dependent|
|Western Blot (WB)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 2 publications below|
The protein encoded by this gene is a member of the MAP kinase family. MAP kinases 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. This kinase is specifically activated by mitogen-activated protein kinase kinase 5. It is involved in the downstream signaling processes of various receptor molecules including receptor type kinases, and G protein-coupled receptors. In response to extracelluar signals, this kinase translocates to cell nucleus, where it regulates gene expression by phosphorylating, and activating different transcription factors. Four alternatively spliced transcript variants of this gene encoding two distinct isoforms have been reported.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
MAP kinase kinase kinase-2 (MEKK2) regulates hypertrophic remodeling of the right ventricle in hypoxia-induced pulmonary hypertension.
44-612G was used in western blot to test if MAPKKK-2 contributes to right ventricle hypertrophy in hypoxia-induced pulmonary hypertension.
|Brown RD,Ambler SK,Li M,Sullivan TM,Henry LN,Crossno JT,Long CS,Garrington TP,Stenmark KR||American journal of physiology. Heart and circulatory physiology (304:H269)||2013|
|Human||Not Cited||Abl-kinase-sensitive levels of ERK5 and its intrinsic basal activity contribute to leukaemia cell survival.||Buschbeck M,Hofbauer S,Di Croce L,Keri G,Ullrich A||EMBO reports (6:63)||2005|
big MAP kinase 1; BMK-1; BMK1 kinase; BNhp-1; BNhp1 kinase; ERK-5; extracellular signal-regulated kinase 5; extracellular-signal-regulated kinase 5; MAP kinase 7; MAPK 7
BMK1; ERK4; ERK5; MAPK7; PRKM7