Phospho-MEK1 (Thr292) Antibody [F18G17]

Application: Reactivity: Human, Mouse, Rat

Lane 1: MDA-MB-231, Lane 2: C6, Lane 3: C6 (phosphatase treated), Lane 4: NIH/3T3

Usage Information

Dilution
WB1:1000 IP1:50

Application
WB, IP

Reactivity
Human, Mouse, Rat

Source
Rabbit Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW
45 kDa

Datasheet & SDS

Biological Description

Specificity
Phospho-MEK1 (Thr292) Antibody [F18G17] detects endogenous levels of total MEK1 protein only when it is phosphorylated at Thr292.

Clone
F18G17

Synonym(s)
Dual specificity mitogen-activated protein kinase kinase 1; MAP kinase kinase 1; MAPKK 1; MAP2K1; MEK1

Background
Phospho‑MEK1 (Thr292) marks a key negative‑regulatory site on the dual‑specificity kinase MEK1, which sits at the core of the Raf–MEK–ERK MAPK cascade that controls cell proliferation, differentiation, and adhesion. MEK1 contains a proline‑rich insert and an activation loop phosphorylated at Ser217 and Ser221 by Raf‑family kinases, and in addition a regulatory segment encompassing Thr292 that lies outside the catalytic core but interfaces with MEK1–ERK2 and MEK1–PAK1 interactions. In active signaling, integrin‑ and growth‑factor‑driven PAK1 phosphorylates MEK1 at Ser298, which promotes MEK1–ERK2 complex formation and facilitates Raf‑1‑mediated activation of MEK1 by enhancing its encounter with upstream Raf and downstream ERK, thereby potentiating ERK‑dependent transcription and cytoskeletal remodeling. ERK‑dependent phosphorylation of MEK1 at Thr292 induces a negative feedback loop: ERK2‑mediated Thr292 phosphorylation reduces MEK1 kinase activity toward ERK1/2, interferes with MEK1 binding to ERK2, and attenuates PAK1‑dependent phosphorylation at Ser298, thereby dampening subsequent Raf‑mediated phosphorylation of the activation‑loop serines and curtailing the magnitude and duration of ERK signaling following cell‑adhesion events and mitogen stimulation. Thr292 phosphorylation functions as a temporal brake that tunes ERK output to mechanical cues and ECM engagement, and dysregulated MEK1 Thr292 phosphorylation has been implicated in perturbed feedback control of ERK in brain and tumor microenvironments.

Background

Phospho‑MEK1 (Thr292) marks a key negative‑regulatory site on the dual‑specificity kinase MEK1, which sits at the core of the Raf–MEK–ERK MAPK cascade that controls cell proliferation, differentiation, and adhesion. MEK1 contains a proline‑rich insert and an activation loop phosphorylated at Ser217 and Ser221 by Raf‑family kinases, and in addition a regulatory segment encompassing Thr292 that lies outside the catalytic core but interfaces with MEK1–ERK2 and MEK1–PAK1 interactions. In active signaling, integrin‑ and growth‑factor‑driven PAK1 phosphorylates MEK1 at Ser298, which promotes MEK1–ERK2 complex formation and facilitates Raf‑1‑mediated activation of MEK1 by enhancing its encounter with upstream Raf and downstream ERK, thereby potentiating ERK‑dependent transcription and cytoskeletal remodeling. ERK‑dependent phosphorylation of MEK1 at Thr292 induces a negative feedback loop: ERK2‑mediated Thr292 phosphorylation reduces MEK1 kinase activity toward ERK1/2, interferes with MEK1 binding to ERK2, and attenuates PAK1‑dependent phosphorylation at Ser298, thereby dampening subsequent Raf‑mediated phosphorylation of the activation‑loop serines and curtailing the magnitude and duration of ERK signaling following cell‑adhesion events and mitogen stimulation. Thr292 phosphorylation functions as a temporal brake that tunes ERK output to mechanical cues and ECM engagement, and dysregulated MEK1 Thr292 phosphorylation has been implicated in perturbed feedback control of ERK in brain and tumor microenvironments.

References
https://pubmed.ncbi.nlm.nih.gov/27342992/ https://pubmed.ncbi.nlm.nih.gov/25971971/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%