The organoids were characterized by immunofluorescence following the protocol of Dekkers et al. (Dekkers et al., 2019 (link)). The following primary antibodies were used to characterize HGOC patients-derived organoids culture in static and passive flow conditions: PAX8 (ProteinTech Group, Germany, EU), WT1 (Abcam, U.K.), and CA-125 (Santacruz Biotechnology, TX). The secondary antibodies that were used are Goat anti-Rabbit IgG (H + L) Alexa Fluor™ Plus 488 (Thermo Fisher Scientific Waltham, Massachusetts, United States) and Goat anti-Mouse IgG (H + L) Alexa Fluor™ Plus 488 (Thermo Fisher Scientific Waltham, Massachusetts, United States). The images were acquired with an EVOS FL Auto 2 (Thermo Fisher Scientific Waltham, Massachusetts, United States).
Ca 125
Manufactured by Santa Cruz Biotechnology
Sourced in United Kingdom
CA-125 is a laboratory assay used to measure the levels of the CA-125 biomarker in a patient's blood sample. CA-125 is a protein that is often elevated in certain types of cancer, particularly ovarian cancer. The assay is designed to quantify the amount of this protein present, providing healthcare professionals with information that may aid in the diagnosis, monitoring, and management of various medical conditions.
Automatically generated - may contain errors
Lab products found in correlation
Ultravision lp detection system hrp dab kit, by Thermo Fisher Scientific (2 mentions)
Evos fl auto 2, by Thermo Fisher Scientific (1 mentions)
Goat anti rabbit igg h l alexa fluor plus 488, by Thermo Fisher Scientific (1 mentions)
Goat anti mouse igg h l alexa fluor plus 488, by Thermo Fisher Scientific (1 mentions)
St5020 multistainer, by Leica (1 mentions)
Ice cold cell recovery solution, by Corning (1 mentions)
Bio agar, by Bio-Optica (1 mentions)
3 protocols using ca 125
1
Immunofluorescence Characterization of HGOC Organoids
2
Characterization of Patient-Derived Tumor Organoids
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Sections of formalin-fixed,
paraffin-embedded ascites and solid tumor organoids were used for
histopathological analyses. Organoids were collected, fixed in phosphate-buffered
10% formalin, and embedded in paraffin using Micro NextGen CelBloking
Kit (Cat no: M20; AV BioInnovation) following manufacturer’s
instructions. Subsequently, 5 μm sections were stained with
hematoxylin and eosin (H&E) using the Leica ST5020 multistainer
and 2 μm sections were cut for IHC analysis. IHC staining was
performed with UltraVision LP Detection System HRP DAB kit (Thermo
Scientific, Waltham). Heat-induced antigen retrieval was performed
using 10 mM citrate buffer pH 6.0. The following antibodies were used
to characterize patient’s derived organoids and parent tumor:
PAX8 (ProteinTech Group, Germany, EU; 10336-1-AP); Ca125 (Santacruz
Biotechnology, TX; sc-52095); WT1 (Abcam, U.K.; ab89901). Tissues
were analyzed with a light microscope using different magnifications.
paraffin-embedded ascites and solid tumor organoids were used for
histopathological analyses. Organoids were collected, fixed in phosphate-buffered
10% formalin, and embedded in paraffin using Micro NextGen CelBloking
Kit (Cat no: M20; AV BioInnovation) following manufacturer’s
instructions. Subsequently, 5 μm sections were stained with
hematoxylin and eosin (H&E) using the Leica ST5020 multistainer
and 2 μm sections were cut for IHC analysis. IHC staining was
performed with UltraVision LP Detection System HRP DAB kit (Thermo
Scientific, Waltham). Heat-induced antigen retrieval was performed
using 10 mM citrate buffer pH 6.0. The following antibodies were used
to characterize patient’s derived organoids and parent tumor:
PAX8 (ProteinTech Group, Germany, EU; 10336-1-AP); Ca125 (Santacruz
Biotechnology, TX; sc-52095); WT1 (Abcam, U.K.; ab89901). Tissues
were analyzed with a light microscope using different magnifications.
3
Histological Analysis of HGOC Organoids
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Sections of formalin-fixed, paraffin-embedded ascites and solid tumor patient-derived organoids were used for histopathological analyses. Organoids were recovered from BME using ice-cold cell recovery solution (Corning, New York, United States) in accordance with manufacturing protocols, fixed in phosphate-buffered 10% formalin, and embedded in 500 μL of Bio-Agar (Bio-Optica Milano Spa, Milano, ITA). Five μm sections were stained with hematoxylin and eosin (H&E) using a Leica ST5020 multistainer and 2 μm sections were cut for IHC analysis. The IHC was performed with an UltraVision LP Detection System HRP DAB kit (Thermo Scientific, Waltham, USA). Heat-induced antigen retrieval was performed using 10 mM citrate buffer pH 6.0. The following antibodies were used to characterize HGOC patients-derived organoids and parental tumors: PAX8 (ProteinTech Group, Germany, EU), WT1 (Abcam, U.K.), and CA-125 (Santacruz Biotechnology, TX).
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