K48-linkage Specific Ubiquitin Rabbit mAb - P0CG47

Core Product Specifications and Parameters

Biological Background: Ubiquitin Function and Localization

• Encoded by the UBB gene, polyubiquitin-B is a precursor protein that is cleaved to generate multiple ubiquitin monomers.
• Ubiquitin exists either free or covalently attached to target proteins via an isopeptide bond, serving as a versatile post-translational modifier.
• Polyubiquitin chains can be formed through different lysine linkages (e.g., K6, K11, K29, K33, K48, K63) or linear linkages, each dictating distinct cellular fates.
• K48-linked polyubiquitin chains primarily target proteins for degradation by the 26S proteasome, a critical pathway for protein quality control and cell cycle regulation.
• K63-linked chains are involved in non-proteolytic processes such as endocytosis, DNA damage responses, and activation of the NF-kappa-B transcription factor.
• Subcellular localization includes the cytoplasm, nucleus, and mitochondrial outer membrane, reflecting its diverse functional roles.
• Ubiquitin itself is subject to post-translational modifications, including phosphorylation and ADP-ribosylation, which can modulate its activity and interactions.
• Free (unanchored) polyubiquitin chains also participate in cellular signaling, such as activation of protein kinases, independent of their conjugation to other proteins.

Experimental Guidance and Technical Tips

• The immunogen is a synthetic peptide designed to specifically recognize the K48-linked polyubiquitin topology; consider using linkage-specific controls to confirm antibody specificity.
• For Western blotting (WB), sample preparation under denaturing and reducing conditions is recommended to detect ubiquitin conjugates; validate performance in your specific sample system.
• In immunofluorescence (IF/ICC), proper fixation and permeabilization protocols should be optimized to preserve ubiquitin signals; include negative controls to assess background.
• For ELISA applications, coating antigens should be carefully selected to mimic the K48-linkage epitope; confirm detection limits with known standards.
• Cross-reactivity is reported for human samples only; if working with other species, verify sequence homology and perform additional validation.

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