RCHY1 Rabbit Monoclonal Antibody for WB and ELISA - Q96PM5

Core Product Specifications and Parameters

Biological Background: RCHY1 Function and Localization

• RCHY1 is an E3 ubiquitin-protein ligase that ubiquitinates multiple targets, including p53/TP53, TP73, HDAC1, and CDKN1B (PubMed:16914734, PubMed:17721809, PubMed:18006823). Related references: PMID:16914734 PMID:17721809 PMID:18006823
• It mediates ubiquitination and degradation of p53/TP53, with a preference for tetrameric p53, impacting cell cycle arrest and apoptosis (PubMed:19043414, PubMed:19483087). Related references: PMID:19043414 PMID:19483087
• Monoubiquitinates the translesion DNA polymerase POLH, regulating DNA damage tolerance (PubMed:21791603). Related references: PMID:21791603
• Participates in ribosome-associated quality control by ubiquitinating stalled nascent chains, facilitating their degradation (PubMed:33909987). Related references: PMID:33909987
• An alternative splicing isoform lacks E3 ubiquitin-protein ligase activity.
• Localizes to the nucleus (including nuclear speckles) and cytoplasm.
• Post-translationally modified by phosphorylation.
• Contains C3HC4-type RING finger and CHY zinc finger domains, which bind zinc and are essential for catalytic function.

Experimental Guidance and Technical Tips

• The immunogen corresponds to amino acids 160-261 of human RCHY1, a region that may provide accessible epitopes under denaturing (WB) and native (ELISA) conditions.
• For WB analysis, load whole-cell lysates from human cell lines and expect a band near 30 kDa; validate with a known positive control and an appropriate negative control (e.g., RCHY1 knockout cells).
• For ELISA, coat recombinant RCHY1 protein as a standard and confirm specificity by competition or by testing the antibody on target-depleted samples.
• Since RCHY1 is a phosphoprotein, consider phosphatase treatment experiments if band shifts are observed.
• Always titrate the antibody from a recommended starting concentration to determine optimal working dilution for each assay.

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