rabbit anti mouse tomm20 proteintech  (Proteintech)

Journal: Scientific Reports

Article Title: Therapeutic potential of human mesenchymal stromal cell-derived mitochondria in a rat model of surgical digestive fistula

doi: 10.1038/s41598-025-13887-3

Figure Lengend Snippet: Characterization of human MSCs-derived mitochondria. ( A ) TOM20 (green) staining and ( Bi – Biii ) MitoTracker red/TOM20 colabling of freshly isolated mitochondria indicating isolation of numerous viable mitochondria from hMSCs. ( C , D ) Scanned electron microscopy (SEM) and transmission electron microscopy using negative staining ( Di , Dii ) as well as cross-sectioned methods ( Ei , Eii ) depicting structurally intact mitochondria of various diameter ranging from 100–1200 nm. ( F , G ) Quality control analysis of isolated mitochondria by Western blotting of mitochondria membrane integrity markers and human OXPHOS complexes ( GI – GV ) showing co-expression of mitochondrial structural and functional proteins in mitochondria preparations (Mito) compared to their corresponding supernatant (SN) loaded with the same amount of protein (Uncropped western blots can be found in the Supplementary Fig. 1). ( H ) ILM of mitochondria samples using Videodrop particle counter showing the size distribution graph with a mean size of 366 nm and a representative interferometry image (depicting the particles in white circles appeared for a moment or the tracked particles shown in circles with orange color). IM inner membrane, OM outer membrane, IMS intermembrane space.

Article Snippet: Initially, isolated mitochondria were incubated either with an Alexa Fluor ® 488 Anti-TOMM20 (translocase of outer mitochondrial membrane 20 homolog) antibody [EPR15581-39] - Mitochondrial Marker (ab205486, abcam) or with a primary rabbit anti-mouse anti-TOM20 antibody (11802-1-AP, Proteintech) for 30 min followed by 30 min incubation in the dark with a secondary antibody (Goat anti-Rabbit IgG) conjugated with green fluorescent AlexaFluor 488 (A-11008, ThermoFisher).

Techniques: Derivative Assay, Staining, Isolation, Electron Microscopy, Transmission Assay, Negative Staining, Control, Western Blot, Membrane, Expressing, Functional Assay

Journal: bioRxiv

Article Title: Rapid and Efficient Quality Control Analysis of Isolated Mitochondria by Interferometric Light Microscopy

doi: 10.1101/2025.05.17.654679

Figure Lengend Snippet: (a) (a) CellInsight™ confocal microscopy image showing fluorescence labeling of isolated mitochondria with TOM20. (b) Quantification of TOM20+ particles (white spots circled in blue, with a higher-resolution view shown in the inset-bi) measured using automated HCS Studio Cell Analysis software. (c) Particle-size distribution of TOM20+ mitochondria preparations. (d-i) Representative TEM images (negative staining) showing an overview and higher magnification of several mitochondria-like structures (e–i). (j) Particle-size distribution for the same samples detected by TEM (negative staining). (k) Graph showing a significantly larger mean diameter measured by IF or for mitochondria-like structures measured by cross-sectioning TEM. No significant differences were found between the mean particle diameters detected by Videodrop or TEM (negative staining or cross-sectioning methods), including all particles or excluding those with diameters outside the detection range for Videodrop (Supplementary Fig. 1). The same experiments (n = 3) were used for different quantification methods in this figure. Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparison tests (k and l). **** p < 0.0001. Error bars represent SEMs. ns: not significant. IF: immunofluorescence microscopy, TEM: transmission electron microscopy, NS: negative staining method, CS: cross-sectioning method.

Article Snippet: Samples of isolated mitochondria were incubated for 30 min with a primary rabbit anti-mouse TOM20 (translocase of outer mitochondrial membrane 20 homolog) antibody (Proteintech), which binds specifically to the TOM20 complex located in the mitochondrial outer membrane.

Techniques: Confocal Microscopy, Fluorescence, Labeling, Isolation, Cell Analysis, Software, Negative Staining, Comparison, Immunofluorescence, Microscopy, Transmission Assay, Electron Microscopy

Journal: Endocrinology

Article Title: Luteal Lipid Droplets: A Novel Platform for Steroid Synthesis

doi: 10.1210/endocr/bqad124

Figure Lengend Snippet: Characteristics of antibodies used for Western blotting and microscopy

Article Snippet: Antibody name Dilution ratio Species specificity Source Supplier (distributor) Catalog No RRID PLIN 2 1:1000 Bovine Guinea pig pAB Fitzgerald Industries International (Acton) 20R-AP002 AB_1282475 PLIN 3 1:1000 Bovine Guinea pig pAB Fitzgerald Industries International 20R-TP001 AB_1289040 STAR 1:10000 Mouse Rabbit pAB Abcam ab96637 AB_10678397 CYP11A1 1:1000 Mouse Rabbit mAB Cell Signaling 14217 AB_2631970 HSD3B 1:1000 a /1:200 b Bovine Mouse mAB Thermo Fisher MA1-46438 AB_2120102 VIM 1:1000 Bovine Rabbit pAB Abcam ab137321 AB_2921312 HSL 1:1000 Mouse Rabbit pAB Cell Signaling 4107 AB_2296900 COX IV 1:1000 Bovine Rabbit mAB Cell Signaling 4850 AB_2085424 HSP47 1:1000 Bovine Mouse mAB Enzo Life Sciences Inc SPA-470 AB_2185189 TOM70 1:1000 Mouse Mouse pAB Abcam ab89624 AB_2043080 CNX 1:200 Mouse Rabbit mAB Cell Signaling 2679 AB_2228381 TOM20 1:200 c Mouse Rabbit mAB Cell Signaling 42406S AB_2687663 TUBB 1:5000 Bovine Mouse mAB Sigma Life Science T4026 AB_477577 ACTB 1:5000 Bovine Mouse mAB Sigma Life Science A5441 AB_476744 Lipi-Blue 1 μM Dojindo Molecular LD01 BODIPY 493/503 10 μM Thermo Fisher D3922 MitoTracker Red FM 200nM Thermo Fisher M22425 HRP-linked 1:10000 Anti-guinea pig Jackson ImmunoResearch 106-035-003 AB_2337402 HRP-linked 1:10000 Anti-rabbit Jackson ImmunoResearch 111-035-003 AB_2313567 HRP-linked 1:10000 Anti-mouse Jackson ImmunoResearch 115-035-205 AB_10015289 DyLight 405 1:500 Anti-mouse Jackson ImmunoResearch 115-475-166 AB_2338786 Alexa Fluor 488 1:500 Anti-mouse Invitrogen A32723 AB_2633275 Alexa Fluor 594 1:500 Anti-rabbit Invitrogen A-11012 AB_2534079 Alexa Fluor 647 1:500 Anti-biotin BioLegend 405237 AB_2941953 Open in a separate window Abbreviations: ACTB, beta-actin (loading control); CNX, calnexin; COX IV, cytochrome c oxidase subunit 4; CYP11A1, cholesterol side-chain cleavage enzyme; HRP, horseradish peroxidase; HSD3B, 3 beta-hydroxysteroid dehydrogenase); HSL, hormone sensitive lipase; HSP47, heat shock protein 47; PLIN2, perilipin 2; PLIN3, perilipin 3; STAR, steroidogenic acute regulatory protein; TOM20, mitochondrial import receptor subunit 20; TOM70, mitochondrial import receptor subunit 70; TUBB, beta-tubulin (loading control); VIM, vimentin. a Dilution used for Western blotting. b Dilution used for confocal microscopy. c Biotinylated antibody.

Techniques: Western Blot

Journal: Endocrinology

Article Title: Luteal Lipid Droplets: A Novel Platform for Steroid Synthesis

doi: 10.1210/endocr/bqad124

Figure Lengend Snippet: Subcellular localization of 3beta-hydroxysteroid dehydrogenase (HSD3B) in steroidogenic small luteal cells. Populations of small luteal cells were enriched from bovine luteal tissue and visualized using confocal microscopy. A, Representative confocal micrograph of HSD3B (red) and luteal lipid droplets (green), calnexin (endoplasmic reticulum; green), or TOM20 (mitochondria; green). White boxes represent an area of 10× enlargement. White dashed lines represent the area of intensity overlap. B, Representative quantitative analysis of intensity overlap. Arrows represent spatial location of lipid droplet. TOM20 (streptavidin, Alexa Fluor 647); calnexin (Alexa Fluor 594); lipid droplet (BODIPY493/503), and HSD3B (DyLight 405). Micron bar represents 20 and 2 µm, respectively.

Article Snippet: Antibody name Dilution ratio Species specificity Source Supplier (distributor) Catalog No RRID PLIN 2 1:1000 Bovine Guinea pig pAB Fitzgerald Industries International (Acton) 20R-AP002 AB_1282475 PLIN 3 1:1000 Bovine Guinea pig pAB Fitzgerald Industries International 20R-TP001 AB_1289040 STAR 1:10000 Mouse Rabbit pAB Abcam ab96637 AB_10678397 CYP11A1 1:1000 Mouse Rabbit mAB Cell Signaling 14217 AB_2631970 HSD3B 1:1000 a /1:200 b Bovine Mouse mAB Thermo Fisher MA1-46438 AB_2120102 VIM 1:1000 Bovine Rabbit pAB Abcam ab137321 AB_2921312 HSL 1:1000 Mouse Rabbit pAB Cell Signaling 4107 AB_2296900 COX IV 1:1000 Bovine Rabbit mAB Cell Signaling 4850 AB_2085424 HSP47 1:1000 Bovine Mouse mAB Enzo Life Sciences Inc SPA-470 AB_2185189 TOM70 1:1000 Mouse Mouse pAB Abcam ab89624 AB_2043080 CNX 1:200 Mouse Rabbit mAB Cell Signaling 2679 AB_2228381 TOM20 1:200 c Mouse Rabbit mAB Cell Signaling 42406S AB_2687663 TUBB 1:5000 Bovine Mouse mAB Sigma Life Science T4026 AB_477577 ACTB 1:5000 Bovine Mouse mAB Sigma Life Science A5441 AB_476744 Lipi-Blue 1 μM Dojindo Molecular LD01 BODIPY 493/503 10 μM Thermo Fisher D3922 MitoTracker Red FM 200nM Thermo Fisher M22425 HRP-linked 1:10000 Anti-guinea pig Jackson ImmunoResearch 106-035-003 AB_2337402 HRP-linked 1:10000 Anti-rabbit Jackson ImmunoResearch 111-035-003 AB_2313567 HRP-linked 1:10000 Anti-mouse Jackson ImmunoResearch 115-035-205 AB_10015289 DyLight 405 1:500 Anti-mouse Jackson ImmunoResearch 115-475-166 AB_2338786 Alexa Fluor 488 1:500 Anti-mouse Invitrogen A32723 AB_2633275 Alexa Fluor 594 1:500 Anti-rabbit Invitrogen A-11012 AB_2534079 Alexa Fluor 647 1:500 Anti-biotin BioLegend 405237 AB_2941953 Open in a separate window Abbreviations: ACTB, beta-actin (loading control); CNX, calnexin; COX IV, cytochrome c oxidase subunit 4; CYP11A1, cholesterol side-chain cleavage enzyme; HRP, horseradish peroxidase; HSD3B, 3 beta-hydroxysteroid dehydrogenase); HSL, hormone sensitive lipase; HSP47, heat shock protein 47; PLIN2, perilipin 2; PLIN3, perilipin 3; STAR, steroidogenic acute regulatory protein; TOM20, mitochondrial import receptor subunit 20; TOM70, mitochondrial import receptor subunit 70; TUBB, beta-tubulin (loading control); VIM, vimentin. a Dilution used for Western blotting. b Dilution used for confocal microscopy. c Biotinylated antibody.

Techniques: Confocal Microscopy

Journal: Endocrinology

Article Title: Luteal Lipid Droplets: A Novel Platform for Steroid Synthesis

doi: 10.1210/endocr/bqad124

Figure Lengend Snippet: Subcellular localization of 3beta-hydroxysteroid dehydrogenase (HSD3B) in steroidogenic large luteal cells. Enriched populations of large luteal cells from bovine luteal tissue were visualized using confocal microscopy. A, Representative confocal micrograph of HSD3B (red) and luteal lipid droplets (green), calnexin (endoplasmic reticulum; green), or TOM20 (mitochondria; green). White boxes represent an area of 10× enlargement. White dashed lines represent the area of intensity overlap. B, Representative quantitative analysis of intensity overlap. Arrows represent spatial location of lipid droplet. TOM20 (streptavidin, Alexa Fluor 647); calnexin (Alexa Fluor 594); lipid droplet (BODIPY493/503), and HSD3B (DyLight 405). Micron bar represents 20 and 2 µm, respectively.

Article Snippet: Antibody name Dilution ratio Species specificity Source Supplier (distributor) Catalog No RRID PLIN 2 1:1000 Bovine Guinea pig pAB Fitzgerald Industries International (Acton) 20R-AP002 AB_1282475 PLIN 3 1:1000 Bovine Guinea pig pAB Fitzgerald Industries International 20R-TP001 AB_1289040 STAR 1:10000 Mouse Rabbit pAB Abcam ab96637 AB_10678397 CYP11A1 1:1000 Mouse Rabbit mAB Cell Signaling 14217 AB_2631970 HSD3B 1:1000 a /1:200 b Bovine Mouse mAB Thermo Fisher MA1-46438 AB_2120102 VIM 1:1000 Bovine Rabbit pAB Abcam ab137321 AB_2921312 HSL 1:1000 Mouse Rabbit pAB Cell Signaling 4107 AB_2296900 COX IV 1:1000 Bovine Rabbit mAB Cell Signaling 4850 AB_2085424 HSP47 1:1000 Bovine Mouse mAB Enzo Life Sciences Inc SPA-470 AB_2185189 TOM70 1:1000 Mouse Mouse pAB Abcam ab89624 AB_2043080 CNX 1:200 Mouse Rabbit mAB Cell Signaling 2679 AB_2228381 TOM20 1:200 c Mouse Rabbit mAB Cell Signaling 42406S AB_2687663 TUBB 1:5000 Bovine Mouse mAB Sigma Life Science T4026 AB_477577 ACTB 1:5000 Bovine Mouse mAB Sigma Life Science A5441 AB_476744 Lipi-Blue 1 μM Dojindo Molecular LD01 BODIPY 493/503 10 μM Thermo Fisher D3922 MitoTracker Red FM 200nM Thermo Fisher M22425 HRP-linked 1:10000 Anti-guinea pig Jackson ImmunoResearch 106-035-003 AB_2337402 HRP-linked 1:10000 Anti-rabbit Jackson ImmunoResearch 111-035-003 AB_2313567 HRP-linked 1:10000 Anti-mouse Jackson ImmunoResearch 115-035-205 AB_10015289 DyLight 405 1:500 Anti-mouse Jackson ImmunoResearch 115-475-166 AB_2338786 Alexa Fluor 488 1:500 Anti-mouse Invitrogen A32723 AB_2633275 Alexa Fluor 594 1:500 Anti-rabbit Invitrogen A-11012 AB_2534079 Alexa Fluor 647 1:500 Anti-biotin BioLegend 405237 AB_2941953 Open in a separate window Abbreviations: ACTB, beta-actin (loading control); CNX, calnexin; COX IV, cytochrome c oxidase subunit 4; CYP11A1, cholesterol side-chain cleavage enzyme; HRP, horseradish peroxidase; HSD3B, 3 beta-hydroxysteroid dehydrogenase); HSL, hormone sensitive lipase; HSP47, heat shock protein 47; PLIN2, perilipin 2; PLIN3, perilipin 3; STAR, steroidogenic acute regulatory protein; TOM20, mitochondrial import receptor subunit 20; TOM70, mitochondrial import receptor subunit 70; TUBB, beta-tubulin (loading control); VIM, vimentin. a Dilution used for Western blotting. b Dilution used for confocal microscopy. c Biotinylated antibody.

Techniques: Confocal Microscopy