Pimonidazole 是一种缺氧标记物，用于瘤内缺氧和细胞增殖的补充性研究。Pimonidazole 通过与大分子的共价结合或通过被还原形成还原性代谢物后在缺氧细胞中积累，可用于定性和定量评估肿瘤缺氧。

Pimonidazole is a novel hypoxia marker for complementary study of tumor hypoxia and cell proliferation in tumor. Pimonidazole accumulates in hypoxic cells via covalent binding with macromolecules or by forming reductive metabolites after reduction of its nitro group, it can be used for qualitative and quantitative assessment of tumor hypoxia .

Solid

Pimonidazole, the exogenous hypoxia marker, is a 2-nitroimidazole compound, which forms covalent bonds with cellular macromolecules at oxygen levels below 1.3%.
Detection: Hypoxic cells were recognized by immunohistochemical detection of pimonidazole using a mouse monoclonal antibody. Cell proliferation was detected with a commercially available monoclonal antibody for proliferating cell nuclear antigen (PCNA). Assessment of hypoxia and cell proliferation was made qualitatively with light microscopy and quantitatively using point counting and image analysis software methods.
The pimonidazole-protein adducts were then detected by immunofluorescence
Cells (90% confluent) are treated with pimonidazole hydrochloride (10-100 μM), diluted in complete media or PBS, and placed in hypoxia or normoxia for 2-4 hours.
Then, cells are washed 4 times in HBSS

Pimonidazole is a 2-nitroimidazole that is reductively activated specifically in hypoxic cells and forms stable adducts with thiol groups in proteins, peptides, and amino acids. Additionally, the amount of pimonidazole that is detected is directly proportional to the level of hypoxia within tumors in vivo.
The following protocol is provided for this useful tool to interrogate the levels of hypoxia in vivo.
1.Pimonidazole hydrochloride is suspended at a concentration of 30 mg/ml in 0.9% sterile saline or PBS.
2.Multiple mice are injected intravenously (tail vein) with 60 mg/kg of the pimonidazole solution.
3.Let circulate in vivo for 90 min before the mice are euthanazied with isoflurane or carbon dioxide (CO2). Note: For animal euthanasia: In compliance with proper IACUC protocols!

Room temperature in continental US; may vary elsewhere.

4°C, sealed storage, away from moistur In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture)

[1]. Varia MA, et al. Pimonidazole: a novel hypoxia marker for complementary study of tumor hypoxia and cell proliferation in cervical carcinoma. Gynecol Oncol. 1998 Nov;71(2):270-7.
[2]. Masaki Y, et al. Imaging Mass Spectrometry Revealed the Accumulation Characteristics of the 2-Nitroimidazole-Based Agent "Pimonidazole" in Hypoxia. PLoS One. 2016 Aug 31;11(8):e0161639.

In Vitro: DMSO : 250 mg/mL (859.85 mM; Need ultrasonic)H₂O : 5 mg/mL (17.20 mM; ultrasonic and warming and heat to 80°C)配制储备液