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- 2025年07月15日
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- 详细信息
- 询价记录
- 文献和实验
- 技术资料
- 英文名：
Recombinant Human Lipocalin-1 Protein, CF
- 规格：
50ug
替代名称
LCN1；lipocalin 1（前白蛋白）；lipocalin 1-like 2；脂质沉积1；Lipocalin-1；MGC71975；PMFA；撕裂脂蛋白；撕裂前白蛋白；薄层色谱；埃布纳腺蛋白；植物蛋白；VEGP；VEGPlipocalin 1（蛋白质迁移速度快于白蛋白、前白蛋白）
Background: Lipocalin-1
Lipocalin-1, also known as tear prealbumin or von Ebner’s gland protein (VEGP), is encoded by the LCN1 gene (1-3). It is a member of the Lipocalin superfamily that binds many different classes of lipophylic chemicals (4). Lipocalin-1 contains three sequence motifs similar to the cystatins, a superfamily of cysteine protease inhibitors (5). In fact, it has been suggested that Lipocalin-1 is a physiological inhibitor of cysteine proteases and plays a role in the control of inflammatory processes in oral and ocular tissues (5). Recombinant Human Lipocalin-1 corresponds to the mature and secreted protein. It is a weak inhibitor of cysteine proteases such as cathepsin V, which is similar to recombinant human Cystatin S (Catalog # 1296-PI).
- Assay Buffer: 50 mM Sodium Acetate, 0.15 M NaCl, pH 5.5
- Cathepsin Buffer: 50 mM Sodium Acetate, 0.15 M NaCl, 10 mM DTT, pH 5.5
- Substrate Buffer: 50 mM Sodium Acetate, 0.15 M NaCl, 5 mM DTT, pH 5.5
- Recombinant Human Lipocalin-1 (rhLipocalin-1) (Catalog # 1708-PI)
- Recombinant Human Cathepsin V (rhCathepsin V) (Catalog # 1080-CY)
- Substrate: Z-Leu-Arg-AMC (Catalog # ES008)
- F16 Black Maxisorp Plate (Nunc, Catalog # 475515)
- Fluorescent Plate Reader (Model: SpectraMax Gemini EM by Molecular Devices) or equivalent
- Prepare a curve of rhLipocalin-1 (MW: 18,805 Da) in Assay Buffer. Make the following serial dilutions: 40,000, 10,000, 4000, 1000, 500, 250, 125, 50, 12.5, and 6.25 nM.
- Dilute rhCathepsin V to 2 µg/mL in Cathepsin Buffer.
- Mix equal volumes of the rhLipocalin-1 curve dilutions and the 2 µg/mL rhCathepsin V. Include a Cathepsin Control (in duplicate) containing Assay Buffer in place of rhLipocalin-1.
- Incubate mixtures at room temperature for 30 minutes.
- Dilute Substrate to 40 µM in Substrate Buffer.
- Load into a black well plate 50 µL of the incubated mixtures, and start the reaction by adding 50 µL of 40 µM Substrate.
- Read at excitation and emission wavelengths of 380 nm and 460 nm (top read), respectively, in kinetic mode for 5 minutes.
- Derive the 50 % inhibiting concentration (IC50) of rhLipocalin-1 by plotting RFU/min (or specific activity) vs. concentration with 4‑PL fitting.
- The specific activity for rhCathepsin V at each point may be determined using the following formula (if needed):
- Redl, B. et al. (1992) J. Biol. Chem. 267:20282.
- Blaker, M. et al. (1993) Biochim. Biophys. Acta 1172:131.
- Lassagne, H. and A.M. Gachon (1993) Exp. Eye Res. 56:605.
- Redl, B. et al. (2000) Biochim. Biophys. Acta 1482:241.
- van’t Hof, W. et al. (1997) J. Biol. Chem. 272:1837.
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