Mouse anti pimonidazole monoclonal antibody

Manufactured by Hypoxyprobe

Sourced in China

The Mouse anti-pimonidazole monoclonal antibody is a laboratory product used for the detection and visualization of hypoxic cells. This antibody specifically binds to pimonidazole adducts formed in cells under low oxygen conditions. It serves as a tool for researchers to study cellular responses to hypoxia.

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Lab products found in correlation

Lsm 710, by Zeiss (2 mentions) Sa00009 1, by Proteintech (1 mentions) S2110, by Solarbio (1 mentions) Fluorescent microscope, by Nikon (1 mentions) Te2000, by Nikon (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) 1 plus kit, by Hypoxyprobe (1 mentions) Biotinylated horse anti mouse igg, by Vector Laboratories (1 mentions) Vp k451, by Abcam (1 mentions) Streptavidin hrp d, by Roche (1 mentions) Tyramide alexa fluor 488, by Thermo Fisher Scientific (1 mentions) Ab28364, by Abcam (1 mentions) Dab map kit, by Roche (1 mentions) Discovery xt processor, by Roche (1 mentions) Cc1 buffer, by Roche (1 mentions)

4 protocols using mouse anti pimonidazole monoclonal antibody

Hypoxia Detection in Murine Colon

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For the detection of hypoxia, mice were treated with 60 mg/kg of pimonidazole HCl (HP1-200Kit; Hypoxyprobe, USA) 30 min prior to sacrifice. Colon samples were fixed in 4% paraformaldehyde, and paraffin-embedded tissues were probed with mouse anti-pimonidazole monoclonal antibody (1:100; Hypoxyprobe) and then stained with Cy-3-conjugated goat anti-mouse antibody (1:50; SA00009-1, Proteintech), and then counterstained with DAPI using anti-fading mountant(S2110, Solarbio, China). Representative images were obtained using a Nikon fluorescent microscope (TE2000, Nikon, Tokyo, Japan).

Ge L., Qi J., Shao B., Ruan Z., Ren Y., Sui S., Wu X., Sun X., Liu S., Li S., Xu C, & Song W. (2022). Microbial hydrogen economy alleviates colitis by reprogramming colonocyte metabolism and reinforcing intestinal barrier. Gut Microbes, 14(1), 2013764.

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Quantifying Tumor Hypoxia and Vasculature

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The variation of hypoxia degree within tumor microenvironment after 24 h post injection of OxgeMCC-r SAE was investigated with immunofluorescence staining. At 90 min before tumors were surgically excised, pimonidazole hydrochloride (Hypoxyprobe-1 plus kit, Hypoxyprobe, USA) was intraperitoneally injected into mice at a dose of 30 mg kg⁻¹. The collected tumor slices were firstly stained with mouse anti-pimonidazole monoclonal antibody (dilution 1:200, Hypoxyprobe) and rat-anti mouse CD31 primary antibody (dilution 1:200, Biolegend) to mark tumor hypoxia regions and blood vessels, respectively. Thereafter, the slices were stained with Alex 488-conjugated goat anti-mouse secondary antibody (Jackson Inc.) and rhodamine-conjugated donkey anti-rat secondary antibody (Jackson Inc.), respectively. The nuclei were stained with 4’,6-diamidino-2-phenylindole (DAPI, Invitrogen). The final images of stained slices were obtained using a confocal laser scanning microscope (Zeiss LSM 710).

Wang D., Wu H., Phua S.Z., Yang G., Qi Lim W., Gu L., Qian C., Wang H., Guo Z., Chen H, & Zhao Y. (2020). Self-assembled single-atom nanozyme for enhanced photodynamic therapy treatment of tumor. Nature Communications, 11, 357.

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Immunohistochemical Analysis of Tumor Hypoxia

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Tumors were fixed in 4% paraformaldehyde, embedded in paraffin, and cut into 5-µm sections. Immunostaining for KIT (Cell Signaling Technology #3074; 1:200), Ki67 (Vector Laboratories #VP-K451; 1:1000), and CD31 (Abcam #ab28364; 1:50) was performed as before (29 (link)). Intratumoral hypoxia was detected by i.p. injection of pimonidazole (60 mg/kg, Hypoxyprobe-1) 1h prior to sacrifice of Kit^V558del/+ mice. Pimonidazole was detected with a Discovery XT processor (Ventana Medical Systems) using deparaffinized tissue sections after antigen retrieval with CC1 buffer (Ventana) and blocking for 30 minutes with Background Buster solution (Innovex). Anti-pimonidazole mouse monoclonal antibody (Hypoxyprobe Inc.) was applied at 0.12 µg/ml and sections were incubated for 5h, followed by 1h incubation with biotinylated horse anti-mouse IgG (Vector Labs, cat# MKB-22258) at 1:200 dilution, then Streptavidin-HRP D (part of DABMap kit, Ventana), and followed by incubation with Tyramide Alexa Fluor 488 (Invitrogen, cat# T20922). Slides were counterstained and analyzed on an Axio2 Imaging wide-field microscope (Zeiss).

Cohen N.A., Zeng S., Seifert A.M., Kim T.S., Sorenson E.C., Greer J.B., Beckman M.J., Santamaria-Barria J.A., Crawley M.H., Green B.L., Rossi F., Besmer P., Antonescu C.R, & DeMatteo R.P. (2015). Pharmacological inhibition of KIT activates MET signaling in gastrointestinal stromal tumors. Cancer research, 75(10), 2061-2070.

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Tumor Hypoxia Imaging in 4T1 Mice

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The 4T1 tumor-bearing BALB/c mice were randomly allocated to four groups: (a) control group (PBS); (b) Au-Pt NPs; (c) X-ray; (d) Au-Pt NPs + X-ray. When the tumor volume reached 60 mm³ at 24 h post-injection of PBS and Au-Pt NPs (20 mg/kg), the mice were exposed to X-ray (8 Gy). The tumor sizes and weight were recorded every other day after treatment. The mice were defined as dead when the tumor volume exceeded 1000 mm³ (Volume = length*width²/2). These mice were sacrificed on day 16 to collect the tumors for the hypoxia staining and crucial organs for the hematoxylin & eosin (H&E) staining.
The tumor tissues were removed for the frozen section. The sections were incubated overnight with an anti-pimonidazole mouse monoclonal antibody (100 times dilution, Hypoxyprobe Inc.). Then, an Alexa Fluo 488 conjugated goat-anti-mouse antibody (100 times dilution, Jackson Inc.) was used as the secondary antibody to combine with the primary antibody. All sections were analyzed and imaged by CLSM (OLYMPUS).

Yang S., Han G., Chen Q., Yu L., Wang P., Zhang Q., Dong J., Zhang W, & Huang J. (2021). Au-Pt Nanoparticle Formulation as a Radiosensitizer for Radiotherapy with Dual Effects. International Journal of Nanomedicine, 16, 239-248.

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