Ubiquitin Rabbit Polyclonal Antibody for WB, IHC-P, IF/ICC, ELISA - P0CG47

Core Product Specifications and Parameters

Biological Background: Ubiquitin Function and Localization

• Ubiquitin is encoded by the UBB gene and produced as polyubiquitin-B, a precursor protein that is processed to yield mature ubiquitin monomers. Ubiquitin is a highly conserved 76-amino acid protein involved in post-translational modification of target proteins.
• Ubiquitin exists either covalently attached to other proteins or free (unanchored). When covalently bound, it is conjugated via an isopeptide bond to target proteins, forming monomers (monoubiquitin) or polyubiquitin chains linked through different Lys residues (Lys-6, Lys-11, Lys-29, Lys-33, Lys-48, Lys-63) or linear chains linked via the initiator Met.
• Different polyubiquitin chain linkages dictate distinct cellular outcomes: Lys-48-linked chains target proteins for proteasomal degradation, while Lys-63-linked chains are involved in endocytosis, DNA damage responses, and NF-kappa-B signaling. Other linkages (Lys-6, Lys-11, Lys-29, Lys-33) participate in DNA repair, ERAD, cell cycle regulation, and kinase modification.
• In addition to conjugated forms, free unanchored polyubiquitin chains also have signaling roles, such as activating protein kinases and modulating other signaling pathways.
• Ubiquitin localizes to multiple subcellular compartments including the cytoplasm, nucleus, and mitochondrion outer membrane, reflecting its broad involvement in cellular processes.
• Post-translational modifications of ubiquitin itself include phosphorylation and ADP-ribosylation, which can modulate its function. The protein contains an isopeptide bond and is part of the Ubl conjugation pathway.

Experimental Guidance and Technical Tips

• The immunogen encompasses full-length ubiquitin (amino acids 1-228), corresponding to the polyubiquitin-B precursor. This polyclonal antibody is expected to recognize both free ubiquitin and ubiquitin-conjugated proteins, but it may also detect polyubiquitin chains. In WB, consider using denaturing and reducing conditions to distinguish monomeric ubiquitin from polyubiquitin conjugates.
• For IHC-P, antigen retrieval may enhance detection, and appropriate controls (e.g., using knockdown or knockout samples) are advisable to confirm specificity. Signal may be diffuse due to ubiquitin's widespread distribution.
• In IF/ICC, fixation and permeabilization methods should be optimized, as ubiquitin localization varies across compartments. Co-staining with organelle markers can aid interpretation.
• ELISA protocols should include standard curves using recombinant ubiquitin to quantify free or total ubiquitin levels. Ensure the antibody is validated for the specific assay format and sample type.
• The antibody shows predicted wide cross-reactivity. For non-human species, validate reactivity in the relevant sample system, as sequence homology is high but may vary.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio offers a one-stop sourcing solution for IVD raw materials, supporting diagnostic manufacturers, research laboratories, and institutes with validated antibody pairs, optimized polyclonal and monoclonal antibodies, bulk ancillary reagents, and rare target raw-material procurement. This ubiquitin antibody supports research in protein degradation, cell signaling, and disease biomarker studies, and CamelBio can assist in scaling up or customizing materials to meet your project needs from concept to clinic.