Immunohistochemistry (IHC) assay was conducted as per the standard protocols. The monoclonal antibodies used for the analyses were as following: rabbit anti-CD66b (Abcam, Cambridge, UK), mouse anti-D2-40 (Dako, Carpinteria, USA), mouse anti-E-cadherin (E-cad) (Dako, Carpinteria, USA), rabbit anti-family with sequence similarity 3 member C (FAM3C) (Abcam, Cambridge, UK), rabbit anti-integrin α6 (Abcam, Cambridge, UK) and rabbit anti-CD151 (Affinity Biosciences, Cincinnati, USA) (for human tissue specimens), mouse anti-E-cad (Cell Signaling Technology, MA, USA), rabbit anti-vimentin (Vim) (Cell Signaling Technology, MA, USA) (for mouse allograft). Integrin α6 (ITGA6) and CD151 were detected using dual-color immunostaining in gastric tumor tissues. The expression intensity of E-cad or Vim was evaluated with mean density (MD) using Image-Pro Plus 6.0 image analysis software (Media Cybernetics, America), and MD means integral optical density (IOD) SUM divided by area SUM.
Mouse anti d2 40
Manufactured by Agilent Technologies
Sourced in United Kingdom
Mouse anti-D2-40 is a primary antibody that recognizes the D2-40 antigen, a podoplanin-specific marker expressed on the surface of lymphatic endothelial cells. It is a useful tool for the identification and enumeration of lymphatic vessels in tissue sections.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 780, by Zeiss (2 mentions)
Mouse anti e cad, by Cell Signaling Technology (1 mentions)
Image pro plus 6.0 image analysis software, by Media Cybernetics (1 mentions)
Rabbit anti von willebrand factor, by Agilent Technologies (1 mentions)
Mouse anti pgp9, by Abcam (1 mentions)
Rabbit anti α smooth muscle actin, by Abcam (1 mentions)
Goat anti mouse alexa fluor 555 antibody, by Thermo Fisher Scientific (1 mentions)
Permafluor, by Thermo Fisher Scientific (1 mentions)
Hoechst 33342, by AnaSpec (1 mentions)
Alexa fluor 488 donkey anti rabbit antibody, by Thermo Fisher Scientific (1 mentions)
2 protocols using mouse anti d2 40
1
Immunohistochemistry Profiling of Gastric Tumor
2
Immunofluorescence Analysis of Airway Tissue
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After the process of deparaffinization and antigen retrieval as described above, tissue sections were permeabilized with 0.1% Tween 20 in PBS. Tissue sections were blocked with blocking reagent and incubated overnight at 4 °C with rabbit anti-α-smooth muscle actin (1:500; Abcam, Cambridge, MA), mouse anti-PGP 9.5 (1:100; Abcam), rabbit anti-von Willebrand factor (1:200; Dako) and mouse anti-D2-40 (1:200; Dako) followed by donkey anti-rabbit Alexa fluor 488 antibody or goat anti-mouse Alexafluor 555 antibody (Invitrogen, Burlington, ONT) for 1 h. Diluent without primary antibodies was used as control. After washing with PBS, cell nucleus was stained with Hoechst 33342 (AnaSpec, Fremont, CA) for 10 min and mounted with PermaFluor (Thermo scientific, Fremont, CA). Images were captured with a confocal microscope (LSM780; Carl Zeiss Microscopy, Jena, Germany) and analyzed using ImageJ (National Institutes of Health, Bethesda, MD) software. ASM and nerve area were expressed as percentage of α-ASMor PGP 9.5-positive area over whole airway area in the section, calculated using ImageJ software.
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