Phospho-MEK1 (Thr292) Monoclonal antibody

Phospho-MEK1 (Thr292) Monoclonal Antibody for WB, FC, ELISA

Host / Isotype

Mouse / IgG1

Reactivity

Human, mouse, rat

Applications

WB, FC, ELISA

Conjugate

Unconjugated

Publications

1

CloneNo.

2D7A8

Cat no : 67873-1-Ig

Synonyms

ERK activator kinase 1, MAP kinase kinase 1, MAP2K1, MAPK/ERK kinase 1, MAPKK 1, MAPKK1, MEK 1, MEK1, MKK1, Phospho-MEK1 (Thr292), PRKMK1

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Validation Data Gallery

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  WB analysis using 67873-1-Ig

  Non-treated A431 and Nocodazole treated A431 cells were subjected to SDS PAGE followed by western blot with 67873-1-Ig (Phospho-MEK1 (Thr292) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with GAPDH antibody as loading control.

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  WB analysis using 67873-1-Ig

  Non-treated HeLa and Calyculin A treated HeLa cells were subjected to SDS PAGE followed by western blot with 67873-1-Ig (Phospho-MEK1 (Thr292) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with GAPDH antibody as loading control.

• 

  WB analysis using 67873-1-Ig

  Non-treated cells and Calyculin A treated cells were subjected to SDS PAGE followed by western blot with 67873-1-Ig (Phospho-MEK1 (Thr292) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with GAPDH antibody as loading control.

• 

  FC experiment of HeLa using 67873-1-Ig

  1X10^6 HeLa cells untreated (dashed lines) or Calyculin A (red) treated were intracellularly stained with 0.13 ug Anti-Human Phospho-MEK1 (Thr292) (67873-1-Ig, Clone:2D7A8) and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) at dilution 1:1000, or 0.13 ug Control Antibody (blue). Cells were fixed with 4% PFA and permeabilized with 90% MeOH.

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Tested Applications

• Positive WB detected in: NIH/3T3 cells, A431 cells, Calyculin A treated HeLa cells, Nocodazole treated A431 cells, Calyculin A treated NIH/3T3 cells, Calyculin A treated HSC-T6 cells
• Positive FC detected in: Calyculin A treated HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:10000
• Flow Cytometry (FC): FC : 0.13 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 1 publications below

Product Information

67873-1-Ig targets Phospho-MEK1 (Thr292) in WB, FC, ELISA applications and shows reactivity with Human, mouse, rat samples.

• Tested Reactivity: Human, mouse, rat
• Cited Reactivity: rat
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: Peptide
• Full Name: mitogen-activated protein kinase kinase 1
• Calculated Molecular Weight: 43 kDa
• Observed Molecular Weight: 40-50 kDa
• GenBank Accession Number: BC139729
• Gene Symbol: MAP2K1
• Gene ID (NCBI): 5604
• RRID: AB_2918631
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

MAP2K1 encodes MAPK1, also known as MEK1. MEK1 variants can enhance MEK1 expression and ERK1 phosphorylation that together lead to continuous activation of MEK/ERK signaling pathway. MEK1 bind directly to ERK2 through a region in the N terminus of MEK. In addition, a proline-rich (PR) regulatory sequence in MEK is also involved in MEK-ERK association and signal propagation. The coupling between MEK1 and ERK2 is enhanced through phosphorylation on S298 in the MEK1 PR region, whereas phosphorylation on MEK1 T292 releases the complex. MEK1 T292 is a substrate of ERK2, but the site is also phosphorylated at a basal level when ERK2 is inhibited, suggesting several regulators of this site . Although the S298 site in MEK2 has been conserved, it lacks the T292 phosphorylation site, and it is not a substrate of PAK1. (PMID: 31972311, PMID: 17928366, PMID: 22177953)

Publications

Species	Application	Title
rat	WB	J Ethnopharmacol; Jia-Wei-Si-Miao-Yong-An Fang stimulates the healing of acute radiation-induced cutaneous wounds through MAPK/ERK pathway; Authors - Yin Wang