4T1 tumor-bearing mice were i.v. injected with PBS or H-MnO2-PEG/C&D. At 6 or 12 h post injection, tumors were surgically excised 90 min after intraperitoneal injection with pimonidazole hydrochloride (60 mg kg−1) (Hypoxyprobe-1 plus kit, Hypoxyprobe Inc.), which was reductively activated in hypoxic cells and formed stable adducts with thiol groups in proteins. For immunofluorescence staining, OCT compound (Sakura Finetek) was employed to prepare frozen sections of the tumors. For detection of pimonidazole, the tumor sections were treated with mouse anti-pimonidazole primary antibody (dilution 1:200, Hypoxyprobe Inc.) and Alex 488-conjugated goat anti-mouse secondary antibody (dilution 1:200, Jackson Inc.) following the kit’s instructions. Tumor blood vessels were stained by rat anti-CD31 mouse monoclonal antibody (dilution 1:200, Biolegend) and Rhodamine-conjugated donkey anti-rat secondary antibody (dilution 1:200, Jackson), subsequently. Cell nuclei were stained with DAPI (dilution 1:5000, Invitrogen). The obtained slices were observed by a confocal microscopy (Leica SP5).
Rat anti cd31 mouse monoclonal antibody
Manufactured by BioLegend
The Rat anti-CD31 mouse monoclonal antibody is a laboratory tool used for the identification and isolation of CD31-positive cells. CD31, also known as PECAM-1 (Platelet Endothelial Cell Adhesion Molecule-1), is a cell surface glycoprotein expressed on endothelial cells, platelets, and some leukocytes. This antibody can be used in various applications such as flow cytometry, immunohistochemistry, and immunoprecipitation to detect and study CD31-expressing cells.
Automatically generated - may contain errors
Lab products found in correlation
1 plus kit, by Hypoxyprobe (3 mentions)
Rhodamine conjugated donkey anti rat secondary antibody, by Jackson ImmunoResearch (3 mentions)
Oct compound, by Sakura Finetek (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Mouse anti pimonidazole primary antibody, by Hypoxyprobe (1 mentions)
Fitc mab1, by Hypoxyprobe (1 mentions)
Mouse anti hif 1α monoclonal antibody, by Abcam (1 mentions)
2 4 amidinophenyl 6 indolecarbamidine dihydrochloride dapi, by Thermo Fisher Scientific (1 mentions)
4 protocols using rat anti cd31 mouse monoclonal antibody
1
Hypoxia Imaging in 4T1 Tumor Mice
2
Tumor xenograft vascular imaging
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Nude mice bearing 4T1 tumors (~200 mm3) were i.v. injected with Cy5.5-HSA or Cy5.5-HSA-PTX. 24 h after injection, tumors were then surgically excised for frozen sections. The blood vessels were stained by incubation with rat anti-CD31 mouse monoclonal antibody (dilution 1:200, Biolegend) and Rhodamine-conjugated donkey anti-rat secondary antibody (dilution 1:200, Jackson).
3
Hypoxia Visualization in 4T1 Tumor-Bearing Mice
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For immunohistochemistry analysis, 4T1 tumor-bearing mice were intravenously injected with TiH1.924-PVP (20 mg·kg−1). At 8 h p.i., tumors on these mice were exposed to the 1064-nm laser irradiation for 20 min with their temperature maintained at ~45 °C. Then immediately, tumors were surgically excised for hypoxia staining assay using the Hypoxyprobe-1 plus kit (Hypoxyprobe Inc) following the standard protocol61 (link),62 . Anti-pimonidazole mouse monoclonal antibody conjugated with FITC (FITC-Mab1, Hypoxyprobe Inc.; Cat. No.: HP2-100Kit; Lot No.: 04-11-19; Clone: 4.3.11.3; Dilution: 1:200) and Alex 488-conjugated goat anti-mouse secondary antibody (Jackon Inc., Cat. No.: 115-545-003, Lot No.: 146108, RRID: AB_2338840; dilution: 1:200) for hypoxia staining. Rat anti-CD31 mouse monoclonal antibody (Biolegend Inc., Cat. No.: 102402, Lot No.: B226360, Clone: 390; dilution: 1:100) and Rhodamine-conjugated donkey anti-rat secondary antibody (Jackon Inc. Cat. No.: 712-025-150, Lot No.: 147079, RRID: AB_2340635; Dilution: 1:200) for blood vessel staining.
4
Evaluating Tumor Hypoxia and Vascularization
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Nude mice bearing 4T1 tumors were i.v. injected with HSA or HSA-PTX (200 μg of PTX, 3 mg of HSA). At 24 h post injection, mice were then i.v. injected with pimonidazole hydrochloride (60 mg/kg) (Hypoxyprobe-1 plus kit, Hypoxyprobe Inc). 90 min later, tumors on those mice were then surgically excised for frozen sections. The tumor slices were incubated with mouse anti-pimonidazole antibody (dilution 1:200, Hypoxyprobe Inc.) and Alex 488-conjugated goat anti-mouse secondary antibody (dilution 1:200, Jackson Inc.) following the kits' instructions. The blood vessels were stained by incubation with rat anti-CD31 mouse monoclonal antibody (dilution 1:200, Biolegend) and Rhodamine-conjugated donkey anti-rat secondary antibody (dilution 1:200, Jackson). Cell nuclei were stained with 2-(4-amidinophenyl)-6-indolecarbamidine dihydrochloride (DAPI) (dilution 1:5000, Invitrogen). The images were captured with a confocal fluorescence microscopy (Leica SP5). For HIF-1α staining, mouse anti-HIF-1α monoclonal antibody (Abcam113642, 200 times dilution) was used as the primary antibody, while goat-anti-mouse IgG antibody conjugated with FITC (Biolegend) was used as the secondary antibody.
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