PLOD3 Rabbit mAb - O60568

Core Product Specifications and Parameters

Biological Background: PLOD3 Function and Localization

• Protein identity and function: PLOD3, also known as LH3 (lysyl hydroxylase 3), is a multifunctional enzyme that performs critical post-translational modifications on collagen lysine residues.
• Hydroxylase activity: It catalyzes the hydroxylation of lysine in -Xaa-Lys-Gly- motifs of procollagens, generating hydroxylysine. This step is essential for collagen thermal stability and cross-link formation. Related references: PMID:11956192, PMID:12475640, PMID:18298658
• Glycosyltransferase activities: Following hydroxylation, the enzyme sequentially adds galactose and glucose to hydroxylysine, forming 1,2-glucosylgalactosyl-5-hydroxylysine. These glycosylation events are indispensable for type IV collagen secretion, basement membrane assembly, and also occur on MBL1 collagen-like domain. Related references: PMID:12475640, PMID:18298658, PMID:18834968, PMID:10934207, PMID:11896059, PMID:11956192, PMID:25419660
• Subcellular localization: PLOD3 resides predominantly in the rough endoplasmic reticulum (ER) lumen and membrane, with a minor pool secreted into the extracellular space. This is consistent with its role in collagen processing within the secretory pathway.
• Tissue distribution: Ubiquitously expressed, with highest levels in heart, placenta, and pancreas, and lower levels in lung, liver, and skeletal muscle. Related references: PMID:9582318, PMID:9724729
• Cofactor requirements: The hydroxylase domain depends on Fe²⁺, 2-oxoglutarate, O₂, and ascorbate (vitamin C), while the glycosyltransferase activities likely require Mn²⁺.
• Structural features: The enzyme is itself a glycoprotein with intramolecular disulfide bonds.

Experimental Guidance and Technical Tips

• Western blotting: Under reducing and denaturing conditions, a specific band at approximately 85 kDa should be observed. Use appropriate protein loading and validate with a relevant cell line or tissue lysate expressing PLOD3.
• Immunohistochemistry (paraffin): Standard antigen retrieval (e.g., citrate buffer, pH 6.0) and blocking steps are recommended. Optimization of primary antibody concentration and detection system is advised.
• ELISA: The antibody can be used in direct or capture ELISA formats. Titration of both coating antigen and antibody concentration should be performed to determine optimal conditions.
• Immunogen considerations: This antibody was raised against a recombinant protein spanning amino acids 25–306 (N-terminal region). It may detect both full-length PLOD3 and potential N-terminal fragments.

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