Anti-HSP47/SERPINH1 Rabbit Polyclonal Antibody for WB, IF/ICC, ELISA - P50454

Core Product Specifications and Parameters

Biological Background: HSP47/SERPINH1 Function and Localization

• HSP47, also known as Serpin H1, is a 47 kDa heat shock protein (colligin) belonging to the serpin superfamily and is encoded by the SERPINH1 gene in humans.
• It functions as a collagen-specific molecular chaperone in the endoplasmic reticulum, essential for proper collagen biosynthesis, folding, and trimerization.
• The protein resides in the endoplasmic reticulum lumen, where it transiently associates with procollagen intermediates to prevent premature aggregation.
• Alternative names include CBP1, CBP2, gp46, AsTP3, PIG14, RA-A47, and SERPINH2, reflecting its discovery in various cellular and pathological contexts.
• Post-translational modifications include acetylation, phosphorylation, and N-glycosylation, which may regulate its activity and stability.
• HSP47 expression is upregulated under stress conditions (e.g., heat shock) and is critical for collagen homeostasis in connective tissues; mutations in SERPINH1 are associated with osteogenesis imperfecta.

Experimental Guidance and Technical Tips

• The immunogen corresponds to amino acids 139–418 of human HSP47/SERPINH1 (NP_001226.2), expressed as a recombinant fusion protein. This region covers a substantial portion of the serpin domain, likely presenting multiple epitopes for polyclonal antibody generation.
• In Western blot (WB), a band around 46 kDa is expected under reducing and denaturing conditions. Validate detection in relevant cell lines or tissues expressing collagen (e.g., fibroblasts, osteoblasts). Use appropriate loading controls.
• For immunofluorescence/immunocytochemistry (IF/ICC), permeabilization (e.g., with Triton X-100 or saponin) is required to access the ER lumen. Expect a reticular staining pattern typical of ER proteins. Co-stain with an ER marker for confirmation.
• In ELISA, antibody specificity and titer should be optimized using recombinant HSP47 or cell lysates. As a polyclonal reagent, lot-to-lot consistency must be verified for quantitative assays.
• Cross-reactivity with mouse HSP47 is confirmed; for other species, sequence homology should be checked before use.

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