Pimonidazole hydrochloride

Name: Pimonidazole hydrochloride
Brand: MedChemExpress
SKU: HY-105129
Rating: 5.0 (9 reviews)

Cat. No.: HY-105129 Purity: 99.98%: Data Sheet
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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 .

For research use only. We do not sell to patients.

CAS No. : 70132-51-3

Size	Stock	Quantity
Solid + Solvent (Highly Recommended)
10 mM * 1 mL in Water ready for reconstitution	In-stock	Estimated Time of Arrival: December 31
Solution
10 mM * 1 mL in Water	In-stock	Estimated Time of Arrival: December 31
Solid
5 mg	In-stock	Estimated Time of Arrival: December 31
10 mg	In-stock	Estimated Time of Arrival: December 31
25 mg	In-stock	Estimated Time of Arrival: December 31
50 mg	In-stock	Estimated Time of Arrival: December 31
100 mg	In-stock	Estimated Time of Arrival: December 31
250 mg	In-stock	Estimated Time of Arrival: December 31
500 mg	Get quote
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Based on 9 publication(s) in Google Scholar

Other Forms of Pimonidazole hydrochloride:

Pimonidazole In-stock
Pimonidazole hydrochloride (Standard) Get quote

9 Publications Citing Use of MCE Pimonidazole hydrochloride

Flow Cytometry

Histological Imaging/Staining

: Pimonidazole hydrochloride purchased from MedChemExpress. Usage Cited in: J Colloid Interface Sci. 2025 Aug 28;702(Pt 1):138870. [Abstract]; Representative fluorescence images of tumor slices from mice bearing 4 T1 tumors stained with Pimonidazole hydrochloride (PIMO) (60 mg/kg; i.p.; 90 min).

: Pimonidazole hydrochloride purchased from MedChemExpress. Usage Cited in: J Colloid Interface Sci. 2025 Dec 3:706:139602.; The hypoxia level in 4T1 cells was quantitatively assessed using Pimonidazole hydrochloride (PIMO) (100 μM; 60 min) as a probe.

: Pimonidazole hydrochloride purchased from MedChemExpress. Usage Cited in: J Colloid Interface Sci. 2025 Dec 3:706:139602.; Representative fluorescence images of tumor slices from 4 T1 tumor-bearing mice after various treatments were stained for hypoxia with Pimonidazole hydrochloride (PIMO) (60 mg/kg; i.p.; 90 min).

: Pimonidazole hydrochloride purchased from MedChemExpress. Usage Cited in: J Colloid Interface Sci. 2024 May:661:908-922. [Abstract]; Fluorescence images of tumor slices from mice bearing 4T1 tumors stained with the hypoxia probe Pimonidazole hydrochloride (PIMO) (60 mg/kg; i.p.; 90 min) after different treatments were shown.

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Purity & Documentation
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Description

Pimonidazole is a novel hypoxia marker for complementary study of tumor hypoxia and cell proliferation in tumor^[1]. 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 ^[2].

In Vitro

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, fixed for 10 min with 10% neutral buffered formalin at room temperature, washed 3 times with tris buffered saline, and mounted with Prolong Gold with DAPI mounting media^[3].
To monitor the intrinsic hypoxic signal, the tissue is removed and the embryos incubated with 400 μM pimonidazole for 2 h before fixation. Immunostaining is performed using antibodies against pimonidazole-protein adducts (Hypoxyprobe)^[4].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

In Vivo

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!
4. Mice are sacrificed and organs are then removed, weighed, snap-frozen, and stored in plastic scintillation vials at -80 °C.
5. The pimonidazole stain can work on paraffin-embedded tissue, frozen tissue, and cell lines. Organs are embedded in OCT, placed in a cryostat, and cut into 10 μm-thick sections. Sections are placed on microscope slides for staining.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Clinical Trial

Molecular Weight

290.75

Formula

C₁₁H₁₉ClN₄O₃

CAS No.

70132-51-3

Appearance

Solid

Color

White to yellow

SMILES

OC(CN1C=CN=C1[N+]([O-])=O)CN2CCCCC2.Cl

Shipping

Room temperature in continental US; may vary elsewhere.

Storage

4°C, sealed storage, away from moisture

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

Solvent & Solubility

In Vitro:

H₂O : 100 mg/mL (343.94 mM; Need ultrasonic)

Preparing
Stock Solutions

Concentration Solvent Mass	1 mg	5 mg	10 mg

1 mM	3.4394 mL	17.1969 mL	34.3938 mL
5 mM	0.6879 mL	3.4394 mL	6.8788 mL
10 mM	0.3439 mL	1.7197 mL	3.4394 mL

View the Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

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Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

Concentration _(start) × Volume _(start) = Concentration _(final) × Volume _(final)

This equation is commonly abbreviated as: C₁V₁ = C₂V₂

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In Vivo Dissolution Calculator

Please enter the basic information of animal experiments:

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Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.

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Working solution concentration: mg/mL

This product has good water solubility, please refer to the measured solubility data in water/PBS/Saline for details.

The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only.If necessary, please contact MedChemExpress (MCE).

Purity & Documentation

Purity: 99.98%

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Data Sheet (277 KB) SDS (392 KB)

COA (250 KB) HNMR (246 KB) RP-HPLC (205 KB) LCMS (92 KB)

Handling Instructions (2659 KB)

References

[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. [Content Brief]

[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. [Content Brief]

[3]. Kristina Y Aguilera, et al. Hypoxia Studies with Pimonidazole in vivo. Bio Protoc [Content Brief]

[4]. Stephanie M Evans, et al. Molecular probes for imaging of hypoxia in the retina. Bioconjug Chem. . 2014 Nov 19;25(11):2030-7. [Content Brief]

[5]. Stephanie M Evans, et al. Molecular probes for imaging of hypoxia in the retina. Bioconjug Chem. . 2014 Nov 19;25(11):2030-7. [Content Brief]

Complete Stock Solution Preparation Table

Optional Solvent	Concentration Solvent Mass	1 mg	5 mg	10 mg	25 mg

H₂O	1 mM	3.4394 mL	17.1969 mL	34.3938 mL	85.9845 mL
	5 mM	0.6879 mL	3.4394 mL	6.8788 mL	17.1969 mL
	10 mM	0.3439 mL	1.7197 mL	3.4394 mL	8.5985 mL
	15 mM	0.2293 mL	1.1465 mL	2.2929 mL	5.7323 mL
	20 mM	0.1720 mL	0.8598 mL	1.7197 mL	4.2992 mL
	25 mM	0.1376 mL	0.6879 mL	1.3758 mL	3.4394 mL
	30 mM	0.1146 mL	0.5732 mL	1.1465 mL	2.8662 mL
	40 mM	0.0860 mL	0.4299 mL	0.8598 mL	2.1496 mL
	50 mM	0.0688 mL	0.3439 mL	0.6879 mL	1.7197 mL
	60 mM	0.0573 mL	0.2866 mL	0.5732 mL	1.4331 mL
	80 mM	0.0430 mL	0.2150 mL	0.4299 mL	1.0748 mL
	100 mM	0.0344 mL	0.1720 mL	0.3439 mL	0.8598 mL

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.