Anti-CYP51A1 Rabbit Monoclonal Antibody for WB, IHC-P, ELISA - Q16850

Core Product Specifications and Parameters

Biological Background: CYP51A1 Function and Localization

• CYP51A1, also known as lanosterol 14-alpha demethylase, belongs to the cytochrome P450 superfamily and is the only cytochrome P450 enzyme conserved across all biological kingdoms (bacteria, fungi, plants, and animals). It is a key enzyme in the sterol biosynthesis pathway responsible for the conversion of lanosterol to cholesterol in mammals.
• Catalytic activity: It catalyzes the three-step oxidative removal of the 14alpha-methyl group (C-32) of sterols such as lanosterol and 24,25-dihydrolanosterol, producing formate and converting them to 4,4-dimethyl-5alpha-cholesta-8,14,24-trien-3beta-ol and 4,4-dimethyl-8,14-cholestadien-3beta-ol, respectively. These are critical intermediates in cholesterol biosynthesis. Related references: PMID:20149798, PMID:8619637, PMID:9559662
• The enzyme is an integral membrane protein localized to the endoplasmic reticulum and microsomal membranes, consistent with its role in lipid metabolism.
• It contains a heme prosthetic group and requires iron as a cofactor, characteristic of monooxygenase activity that utilizes molecular oxygen and NADPH.
• CYP51A1 exhibits broad substrate specificity; besides mammalian sterols, it can demethylate non-mammalian substrates like eburicol, albeit with lower efficiency. Related references: PMID:9559662
• The protein is ubiquitously expressed, with highest levels observed in testis, ovary, adrenal, prostate, liver, kidney, and lung, reflecting the high demand for cholesterol in steroidogenic tissues and membrane synthesis.
• Alternative splicing generates multiple transcript variants, potentially modulating enzyme function or regulation.
• Post-translational modifications include ubiquitin-like (Ubl) conjugation, which may affect protein stability, localization, or activity.

Experimental Guidance and Technical Tips

• The immunogen used is a recombinant protein corresponding to amino acids 400-500 of human CYP51A1, targeting the C-terminal region. This region is likely well-exposed in native and denatured conditions, making the antibody suitable for multiple applications.
• For Western blot (WB) analysis, a band at approximately 57 kDa is expected. Consider using appropriate molecular weight markers and validate specificity using CYP51A1-positive cell lines or tissues (e.g., liver, testis).
• In immunohistochemistry (IHC-P), antigen retrieval may be required. Test on known positive control tissues such as liver or adrenal gland, and include isotype controls to confirm specific staining patterns at the endoplasmic reticulum.
• For ELISA, the antibody can be used as a detection or capture reagent. Pair with a suitable standard, such as the recombinant immunogen or purified CYP51A1 protein, and validate in the relevant sample matrix.
• Given the high sequence homology among human, mouse, and rat CYP51A1, cross-reactivity is expected. However, experimental validation in each species is recommended if used for non-human samples.
• When handling, store at recommended temperatures and avoid repeated freeze-thaw cycles to preserve activity.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio serves as a comprehensive one-stop sourcing partner for IVD raw materials, supporting diagnostic manufacturers, laboratories, and research institutes from concept to clinic. This anti-CYP51A1 rabbit monoclonal antibody addresses research and assay development needs in cholesterol biosynthesis and sterol metabolism. Beyond individual antibodies, CamelBio supplies validated antibody pairs, optimized monoclonal/polyclonal antibodies, bulk ancillary reagents, and facilitates sourcing of rare-target raw materials, streamlining your workflow and accelerating project timelines.