After dewaxing and hybration, the sectioned samples were blocked with 10% FBS. The sections were incubated with primary antibodies at 4°C overnight. The antibodies used were anti-CD7, anti-CD8, anti-CD14, anti-CD15, anti-CD18, anti-CD19 and anti-carcinoembryonic antigen (CEA) (Abcam, Cambridge, MA, USA). The sections were then incubated with Alexa Fluor 488-conjujated anti-mouse/anti-rabbit secondary antibodies (Cell Signaling Technology, Inc., Danvers, MA, USA), for 1 h at room temperature. Nuclei were stained with Hoechst 33342 (Invitrogen Life Technologies).
Anti cd15
Manufactured by Abcam
Sourced in United States
Anti-CD15 is a laboratory reagent used for the detection and identification of the CD15 cell surface antigen. CD15 is a carbohydrate antigen expressed on the surface of granulocytes, monocytes, and some types of cancer cells. This antibody can be used in various immunoassay techniques to study the presence and distribution of CD15-positive cells in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti cd8, by Abcam (4 mentions)
Anti cd19, by Abcam (3 mentions)
Anti cd18, by Abcam (3 mentions)
Anti cd7, by Abcam (3 mentions)
Alexa fluor 488 conjujated anti mouse anti rabbit secondary antibodies, by Cell Signaling Technology (2 mentions)
Hoechst 33342, by Thermo Fisher Scientific (1 mentions)
Fluorescence microscope, by Olympus (1 mentions)
Anti h3cit, by Abcam (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Anti lgr5, by Abcam (1 mentions)
Anti cea, by Abcam (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Anti ly6g, by Abcam (1 mentions)
Anti cd33, by Abcam (1 mentions)
Anti cd3, by Abcam (1 mentions)
Anti cd11b, by Abcam (1 mentions)
Tween 20, by Thermo Fisher Scientific (1 mentions)
Anti β actin, by Cell Signaling Technology (1 mentions)
Anti cox 4, by Cell Signaling Technology (1 mentions)
Anti mouse cd45 fitc, by Thermo Fisher Scientific (1 mentions)
6 protocols using anti cd15
1
Immunofluorescent Staining of Cell Markers
2
Immunofluorescent and Immunohistochemical Analysis of Endometrial/Decidual Tissues
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For immunofluorescent staining, paraffin slides of endometrial/decidual tissues were processed and incubated with primary antibodies, including anti-CD15 (Abcam, USA; 1:1000) and anti-H3Cit (ab5103, USA; 1:1000), before being stained with fluorescent secondary antibodies and DAPI. The images were acquired with a fluorescence microscope (Olympus, Japan), and the number of endometrial/decidual samples with Nets positive was counted. Meanwhile, the paraffin slides were processed and stained with PAD4 antibody (Affinity, USA; 1:200) before being stained with secondary antibodies and chromogen substrate for immunohistochemical staining.
3
Immunophenotyping of Stem Cells
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The SGECs on coverslips were fixed in 4% paraformaldehyde for 10 min, permeabilized with 0.1% Triton X-100 (Sigma) for 20 min at room temperature. Subsequently, 10% FBS was used to block non-specific binding. The SGECs were then incubated with primary antibodies at 4°C overnight. The antibodies that were used were the following: anti-CD7, anti-CD8, anti-CD14, anti-CD15, anti-CD18, anti-CD19, anti-CEA and anti-LGR5 (Abcam). The cells were then incubated with Alexa Fluor 488-conjujated anti-mouse/anti-rabbit secondary antibodies (Cell Signaling Technology, Inc.) for 1 h at room temperature. Nuclei were stained with Hoechst 33342.
4
Multiparametric Immunofluorescence Analysis of Tumor Infiltrates
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Paraffin-embedded sections were de-paraffinized in xylene and rehydrated using alcohol. Citrate buffer were applied to retrieve antigen by incubating the sections by boiling for 10 min in the microwave. Before incubation with primary antibodies at room temperature for an hour, cells were fixed with 4% paraformaldehyde for 10 min and then rinsed with PBS twice and incubated 20 min with fluorescence-conjugated secondary antibodies. The nuclei staining was located with DAPI (Invitrogen) and coverslipped. Immunofluorescent images were first exported and quality by Akoya system. Tumor cryosections were stained with anti-CD3 (clone: D4V8L; cell signaling technical, Mouse), anti-CD8 (clone: D4W2Z; cell signaling technical, Mouse), anti-CD11b (clone: D6X1N; cell signaling technical, Human, Mouse), anti-Ly-6G (clone: E6Z1T; cell signaling technical, Mouse), anti-CD15 (clone: MC480, Human), and anti-CD33 (clone: EPR23051-101, Abcam, Human). Tissues were quantified with Mantra Quantitative Pathology Workstation.
5
Western Blot Analysis of Immune Cell Markers
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Proteins (20 μg/lane) were fractionated by SDS-PAGE and electrotransferred onto polyvinylidene difluoride membranes. The blots were first incubated for 1 h in a blocking buffer consisting of 0.1% Tween-20 (Invitrogen Life Technologies) and 5% non-fat powdered milk, then incubated with a primary antibody at 4°C overnight. The antibodies that were used were as follows: anti-CD7, anti-CD8, anti-CD14, anti-CD15, anti-CD18, anti-CD19 (Abcam) and anti-β-actin (Cell Signaling Technology, Inc.). A horseradish peroxidase-conjugated, goat anti-mouse/anti-rabbit secondary antibody (Zhongshan Jinqiao Biotechnology Co., Ltd., Beijing, China) then was used and antigen-antibody complexes were detected by chemiluminescence using the BeyoECL Plus kit (BiYunTian Biotechnology Research Laboratory, Haimen, China).
6
Multiparametric Flow Cytometry Analysis of Murine Immune Cell Populations
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The following flow antibodies were purchase from Biolegend: Antimouse CD45-Brilliant Violet 421; Antimouse Ly-6G-Alexa Fluor 488; Antimouse Ly-6G-Alexa Fluor 647; Antimouse CD31-Alexa Fluor 647; Antimouse CD31-PE/Cyanine7; Antimouse CD45 FITC; Anti-mouse/human CD11b-Brilliant Violet 421; Antimouse CD184 (CXCR4)-PerCP/Cyanine5.5; Antimouse CD182 (CXCR2)-APC/Cyanine7.
The following flow antibodies were purchase from Invitrogen: Anti-CD144 (VE-cadherin)-eFluor 660; Anti-CD54 (ICAM-1)-PE; Anti-CD11b-APC; Antimouse F4/80-PE.
The following flow antibodies were purchase from BD Biosciences: Antimouse Ly6G-PE; Anti-Mouse CD16/CD32. Anti-Fibrin (Cat#MABS2155) was purchased from Sigma-Aldrich; Anti-SOD2/MnSOD (Cat#ab68155) and Anti-CD15 (Cat#ab135377) were was purchased from Abcam; Antirabbit β-Actin-HRP (Cat#AF5006) was purchased from Beyotime; Antirabbit IgG-HRP (Cat#7074 S), anti-Ly6G (Cat#87048 S) and anti-COX IV (Cat#4844 S) were purchased from Cell Signaling Technology; Anti-TOMM22 (Cat#66562-1-Ig) was purchased from Proteintech; InVivoMAb antimouse Ly6G (Cat#BE0075-1) and InVivoMAb rat IgG2a isotype control (Cat#BE0089) were purchase from Bio X Cell; Donkey antirabbit IgG (H + L)-Cy3 (Cat#711-165-152) and donkey anti-mouse IgG (H + L)-Cy3 (Cat#715-165-150) were purchased from Jackson ImmunoResearch.
The following flow antibodies were purchase from Invitrogen: Anti-CD144 (VE-cadherin)-eFluor 660; Anti-CD54 (ICAM-1)-PE; Anti-CD11b-APC; Antimouse F4/80-PE.
The following flow antibodies were purchase from BD Biosciences: Antimouse Ly6G-PE; Anti-Mouse CD16/CD32. Anti-Fibrin (Cat#MABS2155) was purchased from Sigma-Aldrich; Anti-SOD2/MnSOD (Cat#ab68155) and Anti-CD15 (Cat#ab135377) were was purchased from Abcam; Antirabbit β-Actin-HRP (Cat#AF5006) was purchased from Beyotime; Antirabbit IgG-HRP (Cat#7074 S), anti-Ly6G (Cat#87048 S) and anti-COX IV (Cat#4844 S) were purchased from Cell Signaling Technology; Anti-TOMM22 (Cat#66562-1-Ig) was purchased from Proteintech; InVivoMAb antimouse Ly6G (Cat#BE0075-1) and InVivoMAb rat IgG2a isotype control (Cat#BE0089) were purchase from Bio X Cell; Donkey antirabbit IgG (H + L)-Cy3 (Cat#711-165-152) and donkey anti-mouse IgG (H + L)-Cy3 (Cat#715-165-150) were purchased from Jackson ImmunoResearch.
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