Tissue sections were rinsed in PBS before adding blocking solution [PBS + 10% (v/v) goat serum (Sigma)/1% bovine serum albumin (Fisher Scientific)/0.3% Triton X-100 (Fisher)] for 1 h. Primary antibodies were then diluted in blocking solution and incubated overnight at 4 °C in a humidified slide staining chamber. Lyve1+ and MHCII+ were detected using rat anti-Lyve1 (Clone: ALY7;eBioscience; 1: 500) and rat anti-MHCII (Clone: M5/114.15.2; Novus Biologicals; 1: 500) primary antibodies, paired with rabbit anti-CD68 (polyclonal; Abcam; 1: 1000)/rabbit anti-Iba1 (polyclonal; Wako; 1: 1000), rabbit anti-CD68 (polyclonal; Abcam; 1: 500), rabbit anti-PGP9.5 (Clone: EPR4118; Abcam; 1: 200), rabbit anti-CD206/MRC1 (Clone E6T5J; Cell Signaling; 1: 500), or rabbit anti-CD31 (Clone: SP38; Invitrogen; 1: 50). T cells were detected with rat anti-CD3 (Clone: CD3-12; Abcam; 1: 100). Slides were then rinsed 10 × in PBS. Secondary antibodies (goat anti-rabbit Alexa Fluor-594 with goat anti-rat Alexa Fluor-488, or goat anti-rabbit Alexa Fluor-488 with goat anti-rat Alexa Fluor-594; 1: 500; Invitrogen) in PBS containing DAPI (500 ng/ml; Sigma) were placed on the slides and incubated for 3 h at RT. Sections were again rinsed 10 × in PBS and coverslipped with FluorSave mounting medium (Millipore).
Rabbit anti cd31
Manufactured by Thermo Fisher Scientific
Sourced in United States
Rabbit anti-CD31 is a primary antibody that recognizes the CD31 cell surface antigen, also known as PECAM-1 (Platelet endothelial cell adhesion molecule-1). CD31 is a transmembrane glycoprotein that is predominantly expressed on the surface of endothelial cells and is involved in cell-cell adhesion. This antibody can be used for the detection and identification of endothelial cells in various research applications.
Automatically generated - may contain errors
Lab products found in correlation
Triton x 100, by Thermo Fisher Scientific (1 mentions)
Bovine serum albumin (bsa), by Thermo Fisher Scientific (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Goat serum, by Merck Group (1 mentions)
Rabbit anti cd68, by Abcam (1 mentions)
Fluorsave mounting medium, by Merck Group (1 mentions)
Rabbit anti iba1, by Fujifilm (1 mentions)
Alexa fluor 488 goat anti rat, by Thermo Fisher Scientific (1 mentions)
Rabbit anti pgp9, by Abcam (1 mentions)
Rat anti cd3, by Abcam (1 mentions)
Anti mouse alexa 488, by Thermo Fisher Scientific (1 mentions)
A11029, by Thermo Fisher Scientific (1 mentions)
Mouse anti zo 1, by Thermo Fisher Scientific (1 mentions)
Triton, by Merck Group (1 mentions)
Pa5 16301, by Thermo Fisher Scientific (1 mentions)
Sc 133256, by Santa Cruz Biotechnology (1 mentions)
Ma5 11315, by Thermo Fisher Scientific (1 mentions)
2 protocols using rabbit anti cd31
1
Immunohistochemical Staining of Tissue Sections
2
Immunofluorescent Staining of Tight Junction Proteins
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Cells seeded on Labtek or transwell were washed with PBS, fixed with cold methanol, permeated with 0.01% Triton (Sigma, St. Louis, MO, USA) solution, blocked with 3% BSA and incubated with the following primary antibodies: rabbit anti-CD31
(1:50, PA516301 Invitrogen), mouse anti-GLUT1 (1:100, MA5-11315 Invitrogen), mouse anti-OCLN (1:50, sc-133256 Santa Cruz Biotechnologies), mouse anti-CLN5 (1:20, 35-2500 Invitrogen), mouse anti-ZO1 (1:50, 33-9100 Invitrogen). Subsequently, cells were incubated with secondary antibodies: Alexa-555 anti-rabbit (1:500, A21429 Invitrogen)
was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprint (which this version posted April 14, 2022. ; https://doi.org/10.1101/2022.04.14.488066 doi: bioRxiv preprint and Alexa-488 anti-mouse (1:500, A11029 Invitrogen), respectively. Phalloidin and DAPI staining were used for actin and nuclei visualization, respectively. Images were acquired with a confocal microscope (Carl Zeiss 710) using the image acquisition program Zen (Carl Zeiss). Images were processed and analyzed using ImageJ software (NIH).
(1:50, PA516301 Invitrogen), mouse anti-GLUT1 (1:100, MA5-11315 Invitrogen), mouse anti-OCLN (1:50, sc-133256 Santa Cruz Biotechnologies), mouse anti-CLN5 (1:20, 35-2500 Invitrogen), mouse anti-ZO1 (1:50, 33-9100 Invitrogen). Subsequently, cells were incubated with secondary antibodies: Alexa-555 anti-rabbit (1:500, A21429 Invitrogen)
was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprint (which this version posted April 14, 2022. ; https://doi.org/10.1101/2022.04.14.488066 doi: bioRxiv preprint and Alexa-488 anti-mouse (1:500, A11029 Invitrogen), respectively. Phalloidin and DAPI staining were used for actin and nuclei visualization, respectively. Images were acquired with a confocal microscope (Carl Zeiss 710) using the image acquisition program Zen (Carl Zeiss). Images were processed and analyzed using ImageJ software (NIH).
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