Anti-Phospho-JNK1-T183/Y185 + JNK2-T183/Y185 + JNK3-T221/Y223 Rabbit Monoclonal Antibody for WB, IF-P, IHC-P, ELISA - P45983/P45984/P53779

Core Product Specifications and Parameters

Biological Background: JNK1/2/3 Function and Localization

• Member of the mitogen-activated protein kinase (MAPK) family, also known as stress-activated protein kinases (SAPK), including JNK1 (MAPK8), JNK2 (MAPK9), and JNK3 (MAPK10).
• Activated by dual phosphorylation on a Thr-Pro-Tyr motif (T183/Y185 in JNK1) by upstream kinases MAP2K4/MKK4 and MAP2K7/MKK7 in response to stress stimuli.
• Phosphorylates and regulates transcription factors, notably c-Jun, ATF2, and ELK1, thereby modulating AP-1 transcriptional activity. Related references: PMID:18307971
• Involved in cell proliferation, differentiation, apoptosis, and autophagy by targeting key proteins such as p53/TP53, BAD, BCL2, and BECN1. Related references: PMID:21364637 PMID:21095239 PMID:18570871
• Subcellular localization includes cytoplasm, nucleus, and synapses; translocation to the nucleus occurs upon activation.
• Contributes to inflammatory signaling by phosphorylating NLRP3, promoting inflammasome assembly. Related references: PMID:28943315
• Regulates circadian rhythm through phosphorylation of the CLOCK-BMAL1 heterodimer. Related references: PMID:22441692
• Alternative splicing generates multiple isoforms (e.g., JNK1 alpha and beta) with distinct substrate-binding preferences.

Experimental Guidance and Technical Tips

• Since this antibody recognizes phosphorylation-dependent epitopes, treat samples with phosphatase inhibitors during lysis and avoid prolonged storage at room temperature to preserve phospho-signals.
• For Western blotting, expect bands at approximately 48-52 kDa corresponding to JNK1/2; additional bands for JNK3 may be observed at similar molecular weights; use positive controls such as UV- or anisomycin-treated cells.
• For immunoprecipitation (IF-P), permeabilization and blocking steps are critical; validate staining in relevant stress-stimulated cell lines.
• For immunohistochemistry (IHC-P), consider antigen retrieval with citrate buffer (pH 6.0) and test antibody dilutions in tissues known to exhibit JNK activation, such as those under inflammatory or apoptotic conditions.
• For ELISA, the antibody may be used as a capture or detection reagent; cross-reactivity with non-phosphorylated JNK proteins should be evaluated using appropriate peptide competition assays.

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