Phospho-MAP2K4-T261 Rabbit pAb - P45985

Core Product Specifications and Parameters

Biological Background: MAP2K4 Function and Localization

• MAP2K4 (also known as MEK4, MKK4, SEK1, JNKK1, among other aliases) is a dual specificity protein kinase that functions as a key component of the mitogen-activated protein (MAP) kinase signaling cascade.
• It is an essential kinase in the stress-activated protein kinase/c-Jun N-terminal kinase (SAP/JNK) pathway, directly phosphorylating and activating JNKs (MAPK8/JNK1, MAPK9/JNK2, MAPK10/JNK3) and also the p38 MAPKs (MAPK11, MAPK12, MAPK13, MAPK14).
• MAP2K4 preferentially phosphorylates the tyrosine residue within the Thr-Pro-Tyr motif of JNKs, while MAP2K7 targets the threonine residue; together they synergistically activate JNKs in response to various stimuli.
• This kinase is required for maintaining peripheral lymphoid homeostasis and is involved in mitochondrial death signaling, including cytochrome c release leading to apoptosis.
• Subcellular localization includes both the cytoplasm and nucleus, consistent with its role in signal transduction from cell surface to the nucleus.
• MAP2K4 is widely expressed across tissues, with particularly abundant expression observed in skeletal muscle.
• The protein undergoes several post-translational modifications, including acetylation, methylation, and phosphorylation, which regulate its activity, and is subject to alternative splicing.

Experimental Guidance and Technical Tips

• As a phospho-specific antibody raised against a synthetic phosphorylated peptide around Thr261, it is critical to include phosphatase inhibitors in lysis buffers to preserve the phosphorylation state of endogenous MAP2K4 during sample preparation.
• Validate antibody specificity by treating cell lysates with lambda phosphatase; successful dephosphorylation should abolish the signal if the antibody strictly recognizes the phospho-T261 form.
• For western blotting, the expected molecular weight is approximately 44 kDa; use appropriate gel percentages and transfer conditions for this size range.
• Cross-reactivity is predicted with human, mouse, and rat based on sequence homology of the immunogen region; however, verification in the relevant species is recommended.
• In ELISA applications, consider using this antibody as a capture or detection reagent, and optimize coating and detection conditions empirically with phosphorylated peptide or protein standards.

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