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Hp1 100kit

Manufactured by Hypoxyprobe
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The HP1-100Kit is a laboratory equipment product manufactured by Hypoxyprobe. It is used for the detection and quantification of hypoxic conditions in biological samples.

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Lab products found in correlation

Ab229960, by Abcam (1 mentions) T6397, by Merck Group (1 mentions) Cloner, by STEMCELL (1 mentions) Sb43152, by Merck Group (1 mentions) 233 fb 25 cf, by R&D Systems (1 mentions) Ldn193189, by Merck Group (1 mentions) Axiovision microscope, by Zeiss (1 mentions) Cy3 conjugated donkey anti goat igg, by Wuhan Servicebio Technology (1 mentions) Gb21404, by Wuhan Servicebio Technology (1 mentions) Gb13063, by Wuhan Servicebio Technology (1 mentions) Prolong gold antifade mountant with dapi, by Thermo Fisher Scientific (1 mentions)

4 protocols using hp1 100kit

1

Validating Tumor Oxygenation Conditions

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To confirm the different oxygenation conditions in the tumor groups, mice from each oxic-condition group (n = 4 per group) were injected intravenously with 60 mg/kg body weight of pimonidazole (HP1-100Kit; Hypoxyprobe), 90 minutes before tumor sampling. For normal conditions, injection was followed by 90 minutes of normal air breathing. For carbogen conditions, injection was followed by 90 minutes of carbogen breathing. For clamped conditions, injection was followed by 15 minutes of air breathing followed by 7 minutes of tumor clamping and finally 68 minutes of air breathing. Tumors were then collected, fixed in FineFIX (84-1717-00; Biosystems), embedded in paraffin, and finally cut into 4-µm sections. Tumor hypoxia was validated on tumor sections using mouse antipimonidazole monoclonal antibody (1:50; HP1-100Kit; Hypoxyprobe) incubated for 1 hour at room temperature. The sections were then incubated for 1 hour with a donkey antimouse AF488 secondary antibody (1:250; A21202, Life Technologies). Image acquisition was performed using an upright Zeiss Axiovision microscope.
Leavitt R.J., Almeida A., Grilj V., Montay-Gruel P., Godfroid C., Petit B., Bailat C., Limoli C.L, & Vozenin M.C. (2024). Acute Hypoxia Does Not Alter Tumor Sensitivity to FLASH Radiation Therapy. International journal of radiation oncology, biology, physics, 119(5).
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2

Profiling Glioma Tissue Biomarkers

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The 60 glioma tissues embedded in paraffin were dewaxed and dehydrated with xylene and an ethanol gradient, respectively. Antigen retrieval (10× Citrate Buffer, pH 6.0, Sigma-Aldrich, United States) was performed for specimens and peroxidase blocking (Servicebio, Wuhan, China)) was carried out, followed by incubation overnight at 4°C with primary antibodies including hypoxyprobe pimonidazole (combined with Mab; 1:200; Cat no. HP1-100 kit; Hypoxyprobe, Inc., United States), HOXC6 (1:8000; Cat no. PA5-41479, Thermo Fisher Scientific, United States), MMP9 (1:500; Cat No. 10375-2-AP, Proteintech, Wuhan, China), WT1 (1:200; Cat No. A16319, Abconal, Wuhan, China), SHOX2 (1:200; Cat No. ab229851, Abcam, United States), MYOD1 (1:500; Cat No. 18943-1-AP, Proteintech, Wuhan, China) and HOXA2 (1:200; Cat No. ab229960, Abcam, United States). After three rounds of washing with PBS, the same DAB staining conditions were performed on each specimen after incubation with the secondary antibody (Boster, Wuhan, China). Additionally, antigen-antibody complex signaling within glioma tissues was observed using an orthotopic light microscope.
Ren P., Wang J.Y., Zeng Z.R., Li N.X., Chen H.L., Peng X.G., Bhawal U.K, & Guo W.Z. (2022). A novel hypoxia-driven gene signature that can predict the prognosis and drug resistance of gliomas. Frontiers in Genetics, 13, 976356.
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3

Generation of Oligocortical Spheroids from hESCs

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Human embryonic stem cells (line H7, WiCell) were grown in mTesR1 media and oligocortical spheroids were generated with minor modifications to the protocol previously described (Madhavan et al., 2018 (link)). Briefly, in the first step of generating oligocortical spheroids, CloneR (Stem Cell Technologies, 5889) was used instead of Y-27632 and Dorsomorphin was replaced with 150nM LDN193189 (Sigma, SML0559). Spheroids were treated with 150nM LDN193189 and 10μM SB-43152 (Sigma, S4317) for the first 6 days followed 20ng/ml FGF-2 (R&D Systems, 233-FB-25/CF) and 20ng/ml EGF (R&D Systems, 236-EG-200) from day 7 to 25.
This was followed by 10ng/ml BDNF (R&D Systems, 248-BD) and 20ng/ml NT-3 (R&D Systems, 267-N3) treatment every other day between days 27 and 40. For OPC development and oligodendrocyte differentiation cultures 10ng/ml PDGF-AA (R&D Systems, 221-AA) and 10ng/ml IGF (R&D Systems, 291-GF-200) were added to cultures every other day between days 51 and 60 an 40n/ml T3 (Sigma, T6397) every other day between days 61 and 70. Spheroids were treated every other day with vehicle DMSO or 300nM MEKi between days 70 and 74 and harvested on day 90. Spheroids were treated with 200μM Hypoxyprobe-1 two hours prior to harvesting for IHC (pimonidazole, Hypoxyprobe Inc, Burlington MA, HP1–100Kit).
Allan K.C., Hu L.R., Scavuzzo M.A., Morton A.R., Gevorgyan A.S., Cohn E.F., Clayton B.L., Bederman I.R., Hung S., Bartels C.F., Madhavan M, & Tesar P.J. (2020). Non-Canonical Targets of HIF1a Impair Oligodendrocyte Progenitor Cell Function. Cell stem cell, 28(2), 257-272.e11.
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4

Hypoxia Mapping in 4T1 Tumor Mice

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Peptides were injected iv into 4T1 tumor‐bearing mice and pimonidazole hydrochloride (60 mg/kg; Hypoxyprobe Inc., HP1‐100Kit) was administered into mice via iv injection. Pimonidazole is a non‐toxic 2‐nitroimidazole compound that binds thiol groups to proteins, peptides, and amino acids in hypoxic regions, and could be used as an effective exogenous hypoxia probe.31 Tumors were surgically excised at 1 h after pimonidazole hydrochloride injection. Tumor tissues were paraffin embedded, sectioned, and stained with FITC‐conjugated mouse anti‐pimonidazole protein adducts antibody (Hypoxyprobe Inc., HP1‐100 Kit; 1:100) and goat anti‐mouse CD31 antibody (Servicebio, GB13063; 1:100). Cy3‐conjugated donkey anti‐goat IgG (Servicebio, GB21404; 1:100) was used as a secondary antibody to stain CD31. The tissue samples on slides were mounted with ProLong™ Gold Antifade Mountant with DAPI (Thermo Fisher Scientific, P36935).
Meng F., Liu J., Wei J., Yang J., Zhou C., Yan J, & Liu B. (2022). Tumor‐penetrating peptide internalizing RGD enhances radiotherapy efficacy through reducing tumor hypoxia. Cancer Science, 113(4), 1417-1427.
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