goat anti cd68  (R&D Systems)

Journal: Andrology

Article Title: Human efferent ductules and epididymis display unique cell lineages with motile and primary cilia.

doi: 10.1111/andr.13732

Figure Lengend Snippet: FIGURE 6 Human efferent ductules contain different types of macrophages depending on their shape and location. (A–C and insets) Representative confocal microscopic images showing a cross-section of human efferent ductules stained for macrophage marker CD163 (white), ERα (green), and nucleus (DAPI, blue). Within the epithelium of the efferent ductules, individual CD163-positive and ERα-negative cells are detected at the base of the epithelium (A-B, indicated by arrowheads; C, marked by circles) and within the tubule lumen (A, indicated by arrows). Furthermore, macrophages are observed at the base of the epithelium and extend membrane projections toward the lumen (B-C, marked by asterisks). Dotted lines denote epithelial boundaries. (D, E, and insets) Representative confocal microscopic images showing a cross-section of human efferent ductules stained for macrophage marker CD163 (white), Tight junction protein Zonula occludens (ZO-1, magenta), and nucleus (DAPI, blue). CD163+ macrophages display membrane extensions (arrowheads) oriented towards the lumen. Lu., Lumen; Sp., Spermatozoa. Scale bar = 25 µm.

Article Snippet: A) Primary antibodies Target Origin species Antigen retrieval Dilution Catalog # Company Acetylated Tubulin Mouse EDTA or citrate buffer 1/600 T7451 Sigma-Aldrich CD3 Rabbit EDTA buffer 1/50 A0452 Dako CD45 Mouse Citrate buffer 1/200 555480 BDPharmingen CD68 Mouse EDTA buffer 1/6000 66231-2-Ig Proteintech CD163 Goat EDTA buffer 1/50 AF1607-SP R&D Systems Cytokeratin 5 Chicken EDTA or citrate buffer 1/500 905904 BioLegend Epcam Rabbit EDTA buffer 1/200 21050-1-AP Proteintech ERα Mouse Citrate buffer 1/200 MCA1799T Bio-Rad FoxJ1 Rabbit Citrate buffer 1/500 HPA005714 Sigma-Aldrich Gamma Tubulin Mouse EDTA or citrate buffer 1/600 Ab27074 Abcam p63 Mouse Citrate buffer 1/200 CM163A BiocareMedical PTGS1 (Cox1) Rabbit Citrate buffer 1/300 Ab109025 Abcam V-ATPase B1 Rabbit EDTA buffer 1/100 Reference in 39 Villin Rabbit Citrate buffer 1/200 Ab130751 Abcam Vimentin Mouse Citrate buffer 1/100 Ab20346 Abcam ZO-1 Mouse EDTA buffer 1/100 ZO1-1A12 Invitrogen B) Secondary antibodies Target Origin species Conjugate Dilution Catalog # Company Rabbit IgG (H+L) Goat Alexa Fluor 647 1/400 1/500 1/600 111-605-144 Jackson ImmunoResearch Laboratories Mouse IgG1 Goat Alexa Fluor 488 1/200 1/400 A21121 Invitrogen Mouse IgG2b Goat Alexa Fluor 568 1/600 A21144 Invitrogen Mouse IgM Goat Alexa Fluor 594 1/300 115-585-075 Jackson ImmunoResearch Laboratories Chicken IgY (IgG) (H+L) Donkey Alexa Fluor 647 1/600 703-605-155 Jackson ImmunoResearch Laboratories Mouse IgG2a Goat Alexa Fluor 647 1/500 A21241 Invitrogen Rabbit IgG (H+L) Donkey Alexa Fluor 568 1/500 1/600 A10042 Invitrogen Mouse IgG Donkey Alexa Fluor 647 1/600 715-605-150 Jackson ImmunoResearch Laboratories Mouse IgG Goat Alexa Fluor 568 1/200 1/500 A11031 Invitrogen Goat IgG Rabbit Alexa Fluor 488 1/400 A21222 Invitrogen

Techniques: Staining, Marker, Membrane

Journal: Andrology

Article Title: Human efferent ductules and epididymis display unique cell lineages with motile and primary cilia.

doi: 10.1111/andr.13732

Figure Lengend Snippet: FIGURE 7 Human efferent ductule intraluminal macrophages are positive for CD68. (A-A’’) Representative confocal microscopic images showing an intraluminal macrophage stained for macrophage markers CD68 (green) and CD163 (white). (B and insets) Representative confocal microscopic images showing a cross-section of human efferent ductules stained for macrophage marker CD68 (green), FoxJ1 (magenta), and nucleus (DAPI, blue). Note the presence of sperm nuclei fragments within the macrophages (B’’, arrows) and intraepithelial CD68+ macrophage (arrowhead). (C–E) Representative images obtained by confocal microscopy at high magnification showing CD68+ macrophages with various morphology, size, and organization inside the lumen of human efferent ductules (green). Macrophage and sperm nuclei are visualized with DAPI (blue). Note the presence of sperm nuclei fragments within the macrophages (C, E, arrows). Lu., Lumen; Sp., Spermatozoa. Scale bar = 25 µm.

Article Snippet: A) Primary antibodies Target Origin species Antigen retrieval Dilution Catalog # Company Acetylated Tubulin Mouse EDTA or citrate buffer 1/600 T7451 Sigma-Aldrich CD3 Rabbit EDTA buffer 1/50 A0452 Dako CD45 Mouse Citrate buffer 1/200 555480 BDPharmingen CD68 Mouse EDTA buffer 1/6000 66231-2-Ig Proteintech CD163 Goat EDTA buffer 1/50 AF1607-SP R&D Systems Cytokeratin 5 Chicken EDTA or citrate buffer 1/500 905904 BioLegend Epcam Rabbit EDTA buffer 1/200 21050-1-AP Proteintech ERα Mouse Citrate buffer 1/200 MCA1799T Bio-Rad FoxJ1 Rabbit Citrate buffer 1/500 HPA005714 Sigma-Aldrich Gamma Tubulin Mouse EDTA or citrate buffer 1/600 Ab27074 Abcam p63 Mouse Citrate buffer 1/200 CM163A BiocareMedical PTGS1 (Cox1) Rabbit Citrate buffer 1/300 Ab109025 Abcam V-ATPase B1 Rabbit EDTA buffer 1/100 Reference in 39 Villin Rabbit Citrate buffer 1/200 Ab130751 Abcam Vimentin Mouse Citrate buffer 1/100 Ab20346 Abcam ZO-1 Mouse EDTA buffer 1/100 ZO1-1A12 Invitrogen B) Secondary antibodies Target Origin species Conjugate Dilution Catalog # Company Rabbit IgG (H+L) Goat Alexa Fluor 647 1/400 1/500 1/600 111-605-144 Jackson ImmunoResearch Laboratories Mouse IgG1 Goat Alexa Fluor 488 1/200 1/400 A21121 Invitrogen Mouse IgG2b Goat Alexa Fluor 568 1/600 A21144 Invitrogen Mouse IgM Goat Alexa Fluor 594 1/300 115-585-075 Jackson ImmunoResearch Laboratories Chicken IgY (IgG) (H+L) Donkey Alexa Fluor 647 1/600 703-605-155 Jackson ImmunoResearch Laboratories Mouse IgG2a Goat Alexa Fluor 647 1/500 A21241 Invitrogen Rabbit IgG (H+L) Donkey Alexa Fluor 568 1/500 1/600 A10042 Invitrogen Mouse IgG Donkey Alexa Fluor 647 1/600 715-605-150 Jackson ImmunoResearch Laboratories Mouse IgG Goat Alexa Fluor 568 1/200 1/500 A11031 Invitrogen Goat IgG Rabbit Alexa Fluor 488 1/400 A21222 Invitrogen

Techniques: Staining, Marker, Confocal Microscopy