Two months after transplantation, sections prepared from the testicular tissue were deparaffinized and dehydrated; then, the sections were boiled in buffer citrate at 100 °C (20 min) for antigen retrieval. In order to block nonspecific antigens and ensure permeabilization, the sections were exposed to 0.1% bovine serum albumin (BSA, Sigma-Aldrich, USA) in 0.1% triton X-100 for 1 h. Then, the sections were incubated at 4 °C overnight with the primary antibodies against the following markers (PLZF, SCP3): anti-PLZF (1:500, Santa Cruz, USA) and anti-SCP3 (1:400, Abcam, USA). Next, the sections on the slides were washed with PBS, and goat anti-rabbit secondary antibody conjugated with FITC (1:400, Abcam, USA) was added to the slides and maintained at room temperature for 2 h. Tissue sections of the testis were stained with DAPI to detect the cell nucleus and then observed under a fluorescence microscope (Olympus LX71, Japan) equipped with a camera.
Anti scp3
Manufactured by Abcam
Sourced in United States
Anti-SCP3 is a laboratory reagent used for the detection and quantification of the SCP3 protein. It is a highly specific antibody that binds to the SCP3 protein, allowing researchers to study its expression and localization in various biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti zpc, by Santa Cruz Biotechnology (2 mentions)
Anti gdf9, by Merck Group (2 mentions)
Anti ssea4, by Merck Group (2 mentions)
Anti tra 1 60, by Merck Group (2 mentions)
Anti tra 1 81, by Merck Group (2 mentions)
Lsm 710, by Zeiss (2 mentions)
Anti plzf, by Santa Cruz Biotechnology (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Dmi3000 microscope, by Leica (1 mentions)
Anti nanog, by Merck Group (1 mentions)
Fluorescein isothiocyanate labeled igg, by Santa Cruz Biotechnology (1 mentions)
Anti oct4, by Santa Cruz Biotechnology (1 mentions)
Anti zo 1, by Abcam (1 mentions)
Anti gata1, by Abcam (1 mentions)
Anti ftl, by Abcam (1 mentions)
Anti ki67, by Abcam (1 mentions)
Anti nanog, by Abcam (1 mentions)
Anti epcam, by Santa Cruz Biotechnology (1 mentions)
Sp8 confocal microscope, by Leica (1 mentions)
4 protocols using anti scp3
1
Immunohistochemical Analysis of Testicular Tissue
2
Immunocytochemical Characterization of Stem Cells
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Epithelial cells, oocyte-like cells, and colonies maintained on hAECs were fixed with 4% paraformaldehyde for 15 to 20 minutes at room temperature and permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature. Cells were then blocked with blocking solution for 30 minutes and incubated with anti-OCT4 (rabbit anti-human 1:200; Santa Cruz), anti-NANOG (rabbit anti-human, 1:200; Chemicon, Rolling Meadows, IL, USA), anti-DAZL (goat anti human, 1:500; Santa Cruz), anti-STELLA (goat anti-human, 1:200; Santa Cruz), anti-ZPC (rabbit anti-human, 1:200; Santa Cruz), anti-SCP3 (rabbit anti-human, 1:800; Abcam), anti-GDF9 (rabbit anti-human, 1:200; Millipore), anti-SSEA4 (mouse anti-human, 1:100; Millipore), anti Tra-1-60 (mouse anti-human, 1:100; Millipore), and anti Tra-1-81(mouse anti-human, 1:100; Millipore) antibody for 1 hour at room temperature. Cells were then probed with fluorescein isothiocyanate-labeled IgG (1:200; Santa Cruz) or Rodamine (TRITC)-labeled IgG (1:100; Invitrogen) and incubated at room temperature for another 20 minutes. The slides were then covered with mounting medium (glycerol diluted 3:1 in PBS; Vector Laboratories, Burlingame, CA, USA). Fluorescence images were obtained with a Leica DMI3000 microscope.
3
Comprehensive Immunofluorescence Analysis of Mouse Testis
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The mice were anesthetized using the 0.4% pentobarbital sodium (1 mL/100 g) solution. After that, the mice were perfused with 0.9% saline solution and treated with 4% paraformaldehyde. The testis and epididymis were carefully dissected and immersed in 30% sucrose solution for 2 days. The testis and epididymis were sliced into 15-μm-thick slices, and then the slices were washed with 0.01 M PBS three times for 5 min each time. The slices were then incubated with goat serum at 37°C for 60 min. Mouse monoclonal FtH antibody (1:200; Cat. No. ab104224, Abcam, Waltham, MA, United States), anti-FtL (1:400; Cat. No. 20403-1-AP ProteinTech, Rosemont, IL, United States), anti-DDX4 (1:400; Cat. No. CL488-67147, ProteinTech, Wuhan, China), anti-SCP3(1:400; No. ab97672, Abcam, Waltham, MA, United States), anti-GATA1 (1:400; Cat. No. 60011-1-Ig, ProteinTech, Wuhan, China), anti-Ki67 (1:400; No. ab115730 Abcam, Waltham, MA, United States), and anti-ZO1 (1:400; Cat. No. ab24950, Abcam, Waltham, MA, United States) were used as primary antibodies. The slices were incubated overnight at 4°C. Next, the slices were incubated with FITC-conjugated and rhodamine-conjugated secondary antibodies. Images were observed using a ZEISS LSM710.
4
Immunophenotyping of Oocyte-like Cells
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IOSE80 cells, oocyte-like cells, and colonies maintained on hAECs were fixed with 4% paraformaldehyde for 15‒20 min at room temperature and permeabilized with 0.1% Triton X-100 for 10 min at room temperature. Cells were then blocked with 5% FBS in PBS for 30 min and incubated with primary antibodies, including anti-NANOG (rabbit anti-human; 1:200; Abcam, Cambridge, UK), anti-EpCAM (goat anti-human; 1:500; Santa Cruz Biotechnology), anti-IFITM3 (rabbit anti-human; 1:200; Novus Biologicals, Littleton, USA), anti-ZPC (rabbit anti-human; 1:200; Santa Cruz Biotechnology), anti-SCP3 (rabbit anti-human; 1:800; Abcam), anti-GDF9 (rabbit anti-human; 1:200; Millipore, Billerica, USA), anti-SSEA4 (mouse anti-human; 1:100; Millipore), anti-Tra-160 (mouse anti-human; 1:100; Millipore), and anti-Tra-1-81 (mouse anti-human; 1:100; Millipore) antibodies for 1 h at room temperature. Cells were then probed with fluorescein isothiocyanate-labelled IgG (1:200; Santa Cruz Biotechnology) or rhodamine (TRITC)-labelled IgG (1:100; Invitrogen, Carlsbad, USA) antibodies and incubated at room temperature for an additional 30 min. Each antibody was detected using corresponding secondary antibodies conjugated to fluorescein isothiocyanate. Fluorescence images were obtained using a Leica DMI3000 microscope or an SP8 confocal microscope (Leica Microsystems, Wetzlar, Germany).
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