guinea pig anti mouse nephrin  (Progen Biotechnik)

Journal: Methods and Protocols

Article Title: Parsing Glomerular and Tubular Structure Variability in High-Throughput Kidney Organoid Culture

doi: 10.3390/mps8050125

Figure Lengend Snippet: Morphology of glomerular and tubular structures in iPSC-derived kidney organoids generated with the original APEL/air-medium interface protocol. Representative immunofluorescence images of the maturing glomerular structures, stained for nephrin, and maturing tubular structures, stained for epithelial cadherin (ECAD) in healthy-donor iPSC lines HEL24.3 and HEL61.2, and the GRACILE iPSC line HEL124.2. Images acquired with ( A ) widefield fluorescence microscopy, scale bars 500 µm, and ( B ) the Opera Phenix spinning disk confocal microscope, scale bars 100 µm.

Article Snippet: Fluorescein Lotus tetragonolobus lectin (LTL; Vector Laboratories, Newark, CA, USA, FL-1321, 1:500) and following primary antibodies were utilized: guinea pig anti-mouse nephrin (Progen, Heidelberg, Germany, GP-N2, 1:300) and mouse anti-human epithelial cadherin (ECAD; BD Biosciences 610181, 1:400).

Techniques: Derivative Assay, Generated, Immunofluorescence, Staining, Fluorescence, Microscopy

Journal: Methods and Protocols

Article Title: Parsing Glomerular and Tubular Structure Variability in High-Throughput Kidney Organoid Culture

doi: 10.3390/mps8050125

Figure Lengend Snippet: Opera Phenix high-content screening and quantification of nephrin and ECAD-positive maturing structures in kidney organoids generated with higher throughput approaches. Four modified high-throughput approaches consisting of different culture systems (monolayer culture, ML, spheroid suspension culture, S, and/or air-medium interface, AMI) and variable amounts of cells at the initiation of spheroids, from 2000 (2 k) to 200,000 (200 k) cells, were tested. Two iPSC lines were utilized (HEL24.3 and HEL61.2) in two different experiments ( A , B ). The proportions of nephrin and ECAD-positive area per organoid (imaged with Opera Phenix) are shown with blue dots (mean ± s.d. for each condition indicated with black crossbars). Unadjusted significance levels are shown (Mann-Whitney U test *: p < 0.05, **: p < 0.01, ns: non-significant). Number of replicate samples per condition provided in .

Article Snippet: Fluorescein Lotus tetragonolobus lectin (LTL; Vector Laboratories, Newark, CA, USA, FL-1321, 1:500) and following primary antibodies were utilized: guinea pig anti-mouse nephrin (Progen, Heidelberg, Germany, GP-N2, 1:300) and mouse anti-human epithelial cadherin (ECAD; BD Biosciences 610181, 1:400).

Techniques: High Content Screening, Generated, Modification, High Throughput Screening Assay, Suspension, MANN-WHITNEY

Journal: Methods and Protocols

Article Title: Parsing Glomerular and Tubular Structure Variability in High-Throughput Kidney Organoid Culture

doi: 10.3390/mps8050125

Figure Lengend Snippet: Representative Opera Phenix immunofluorescence images of nephrin and ECAD-positive maturing structures in kidney organoids generated with the higher throughput approaches. Kidney organoids generated with high-throughput approaches one (S+AMI, a spheroid suspension culture followed by air-medium interface) and four (S, a spheroid suspension culture). Images represent two iPSC lines HEL24.3 and HEL61.2 and two separate experiments ( A , B ) with 50,000 cells utilized for the initiation of spheroids. Organoids were stained for nephrin (green) and ECAD (red) for Opera Phenix high-content screening. Nuclei stained with Hoechst (blue). Scale bar 200 µm. (S) and 500 µm (S+AMI).

Article Snippet: Fluorescein Lotus tetragonolobus lectin (LTL; Vector Laboratories, Newark, CA, USA, FL-1321, 1:500) and following primary antibodies were utilized: guinea pig anti-mouse nephrin (Progen, Heidelberg, Germany, GP-N2, 1:300) and mouse anti-human epithelial cadherin (ECAD; BD Biosciences 610181, 1:400).

Techniques: Immunofluorescence, Generated, High Throughput Screening Assay, Suspension, Staining, High Content Screening

Journal: bioRxiv

Article Title: Parsing glomerular and tubular structure variability in high-throughput kidney organoid culture

doi: 10.1101/2025.05.05.652160

Figure Lengend Snippet: Morphology of renal structures in kidney organoids generated with the original APEL/air-medium interface protocol. Representative immunofluorescence images of the maturing glomerular structures, stained for nephrin, and maturing tubular structures, stained for epithelial cadherin (ECAD) in healthy-donor iPSC lines HEL24.3 and HEL61.2, and the GRACILE iPSC line HEL124.2, images acquired with (A) widefield fluorescence microscopy, scale bar 500µm, and (B) the Opera Phenix spinning disk confocal microscope, scale bar 200µm.

Article Snippet: Fluorescein Lotus tetragonolobus lectin (LTL; Vector Laboratories FL-1321, 1:500) and following primary antibodies were utilized: guinea pig anti-mouse nephrin (Progen GP-N2, 1:300) and mouse anti-human epithelial cadherin (ECAD; BD Biosciences 610181, 1:400).

Techniques: Generated, Immunofluorescence, Staining, Fluorescence, Microscopy

Journal: bioRxiv

Article Title: Parsing glomerular and tubular structure variability in high-throughput kidney organoid culture

doi: 10.1101/2025.05.05.652160

Figure Lengend Snippet: Four modified high-throughput approaches consisting of different culture systems (monolayer culture, ML, spheroid suspension culture, S, and/or air-medium interface, AMI) and variable amounts of cells at the initiation of spheroids, from 2 000 (2k) to 200 000 (200k) cells, were tested. Two iPSC lines were utilized (HEL24.3 and HEL61.2) in two different experiments (A and B). Proportion of nephrin and ECAD-positive area per organoid (imaged with Opera Phenix) is shown with a dot (mean ± s.d. for each condition indicated with black crossbars). Unadjusted significance levels are shown (Mann-Whitney U test *: p < 0.05, **: p < 0.01, ns: non-significant). Number of replicate samples per condition provided in Supplementary Table S1.

Article Snippet: Fluorescein Lotus tetragonolobus lectin (LTL; Vector Laboratories FL-1321, 1:500) and following primary antibodies were utilized: guinea pig anti-mouse nephrin (Progen GP-N2, 1:300) and mouse anti-human epithelial cadherin (ECAD; BD Biosciences 610181, 1:400).

Techniques: Modification, High Throughput Screening Assay, Suspension, MANN-WHITNEY

Journal: bioRxiv

Article Title: Parsing glomerular and tubular structure variability in high-throughput kidney organoid culture

doi: 10.1101/2025.05.05.652160

Figure Lengend Snippet: Kidney organoids generated with the high-throughput approaches 1 (S+AMI, a spheroid suspension culture followed by air-medium interface) and 4 (S, a spheroid suspension culture). Images represent two iPSC lines HEL24.3 and HEL61.2 and two separate experiments (A and B) with 50 000 cells utilized for the initiation of spheroids. Organoids were stained for nephrin (green) and ECAD (red) for Opera Phenix high-content screening. Scale bar 200 µm. (S) and 500 µm (S+AMI).

Article Snippet: Fluorescein Lotus tetragonolobus lectin (LTL; Vector Laboratories FL-1321, 1:500) and following primary antibodies were utilized: guinea pig anti-mouse nephrin (Progen GP-N2, 1:300) and mouse anti-human epithelial cadherin (ECAD; BD Biosciences 610181, 1:400).

Techniques: Generated, High Throughput Screening Assay, Suspension, Staining, High Content Screening

Journal: bioRxiv

Article Title: Parsing glomerular and tubular structure variability in high-throughput kidney organoid culture

doi: 10.1101/2025.05.05.652160

Figure Lengend Snippet:

Article Snippet: Fluorescein Lotus tetragonolobus lectin (LTL; Vector Laboratories FL-1321, 1:500) and following primary antibodies were utilized: guinea pig anti-mouse nephrin (Progen GP-N2, 1:300) and mouse anti-human epithelial cadherin (ECAD; BD Biosciences 610181, 1:400).

Techniques: