Frozen sections of tendon from a patient with RA and cartilage from a patient with osteoarthritis were purchased from OriGene Technologies. The sections were blocked with 2% fetal bovine serum (FBS) in PBS for overnight at room temperature. The sections were incubated with primary antibodies for 3 hours at room temperature. For tendon sections, either rat α-TNF (50 μg/ml) or equimolar CBP–α-TNF, mouse anti-human CD31 antibody (5 μg/ml; Abcam), and rabbit anti-human type I collagen antibody (5 μg/ml; Abcam) were used as primary antibodies. For cartilage sections, either rat α-TNF or equimolar CBP–α-TNF (50 μg/ml) and rabbit anti-human type II collagen antibody (5 μg/ml; Abcam) were used as primary antibodies. After incubating with the fluorescently tagged secondary antibodies, slides were covered with ProLong Gold Antifade Mountant with 4′,6-diamidino-2-phenylindole (Thermo Fisher Scientific). The images were scanned with a Pannoramic Digital Slide Scanner (3DHISTECH) and analyzed using Pannoramic Viewer software (3DHISTECH).
Mouse anti human cd31 antibody
Manufactured by Abcam
Sourced in United States
Mouse anti-human CD31 antibody is a primary antibody that specifically recognizes the CD31 antigen, also known as PECAM-1 (Platelet Endothelial Cell Adhesion Molecule-1). CD31 is a transmembrane glycoprotein expressed on the surface of endothelial cells, platelets, and certain leukocytes.
Automatically generated - may contain errors
Lab products found in correlation
Prolong gold antifade mountant with 4 6 diamidino 2 phenylindole, by Thermo Fisher Scientific (1 mentions)
Pannoramic digital slide scanner, by 3DHISTECH (1 mentions)
Pannoramic viewer software, by 3DHISTECH (1 mentions)
Goat anti human 4igb7 h3 antibody, by R&D Systems (1 mentions)
Rabbit anti human vegfa antibody, by Proteintech (1 mentions)
Rat anti mouse cd31 antibody, by BioLegend (1 mentions)
Goat anti rat igg, by Abcam (1 mentions)
Goat anti mouse igg, by Abcam (1 mentions)
Application suite x, by Leica (1 mentions)
Alexa fluor 488 goat anti rabbit igg secondary antibody, by Abcam (1 mentions)
Normal serum block, by BioLegend (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Abcam (1 mentions)
4 protocols using mouse anti human cd31 antibody
1
Immunohistochemical Analysis of Inflammatory Markers
2
Immunohistochemical Analysis of CRC Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A total of 125 pairs of CRC samples and adjacent normal tissues were obtained from CRC patients at the First Affiliated Hospital of Soochow University (Suzhou, China). All experiments involving patient specimens were approved by the Institutional Review Board of Soochow University. Informed consent was obtained from each patient. Detailed clinicopathological information is provided in Supplementary Table S1 . An IHC assay was conducted as previously described17 (link). Briefly, CRC samples or xenograft tumor samples were deparaffinized and rehydrated. After antigen retrieval with 10 mM sodium citrate buffer (pH 6.0), the sections were incubated with goat anti-human 4IgB7-H3 antibody (R&D Systems, MN, USA, #AF1027, 1:100), mouse anti-human CD31 antibody (Abcam, Cambridge, MA, USA, #ab32457, 1:1500), or rabbit anti-human VEGFA antibody (Proteintech, Wuhan, China, #19003–1-AP, 1:500). All sections were then reviewed blindly by two experienced pathologists (Dr. Cao and Dr. Zhan). The scoring criteria for IHC staining were based on the intensity of immunostaining and the percentage of immunoreactive cells as described previously17 (link). We calculated the score of the MVD on the basis of previously described criteria22 (link).
3
Multicolor Immunostaining of Tumor Vasculature
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The primary antibodies used here included rat anti-mouse CD31 antibody (Biolegend, CA, USA), mouse anti-human CD31 antibody and rabbit anti-human PDGFRβ antibody (Abcam CA, USA). The secondary antibodies were goat anti-rat IgG (DyLight 550), donkey anti-rabbit IgG antibody (DyLight 488 or DyLight 550), and goat anti-mouse IgG (DyLight 550) (Abcam, CA, USA). Tumor tissues derived from patients or mice were frozen, sectioned and fixed with 2 % formaldehyde for 4 min. Subsequently, the tissues were incubated with primary antibody at 37°C for 1.5 h followed by incubation with the corresponding secondary antibody at 37°C for 0.5 h. In addition, the tissues were washed with PBS 4 times prior to incubation with either primary antibody or secondary antibody. The nuclei of cells were visualized using DAPI staining.
To localize the recombinant proteins on PDGFRβ-expressing cells in tumors, FAM-labeled proteins were intravenously injected into mice bearing LS174T tumor xenografts. Subsequently, the tumor xenografts were removed at 5, 30, and 60 min post-injection and frozen and sectioned followed by staining with anti-PDGFRβ antibody or anti-CD31 antibody.
To localize the recombinant proteins on PDGFRβ-expressing cells in tumors, FAM-labeled proteins were intravenously injected into mice bearing LS174T tumor xenografts. Subsequently, the tumor xenografts were removed at 5, 30, and 60 min post-injection and frozen and sectioned followed by staining with anti-PDGFRβ antibody or anti-CD31 antibody.
4
Immunofluorescence Staining of Endothelial Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were rinsed with phosphate-buffered saline (with calcium chloride and magnesium chloride; PBS+/+), fixed with ice-cold methanol for 10 min at − 20 °C, and blocked with 3% (w/v) BSA in PBS−/− for 30 min prior to being stained overnight at 4 °C with 5 µg/mL of primary antibodies in the same blocking solution. The primary antibodies were sourced from Abcam, and included a mouse anti-human CD31 antibody (catalogue no. ab24590), a rabbit anti-human VE-cadherin antibody (catalogue no. ab33168), and a mouse anti-human vWF antibody (catalogue no. ab194405). Cells were then blocked with Normal Serum Block (BioLegend) for 30 min at 25 °C and stained for 1 hr with 2.5 µg/mL of either a goat anti-mouse IgG-Alexa Fluor 594 secondary antibody (BioLegend; catalogue no. 405326) or a goat anti-rabbit IgG-Alexa Fluor 488 secondary antibody (Abcam; catalogue no. ab150077) diluted in 3% (w/v) BSA in PBS−/−. Cells were then stained for 5 min with 3 µM 4′,6-diamidino-2-phenylindole (Abcam) in PBS−/−. Wide-field immunofluorescence microscopy was performed using a Leica DMi8, operated using the Leica Application Suite X software version 3.5.5.19976 (Leica Microsystems). Image processing was performed using Fiji software version 2.1.0/1.53c46 (link).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!