Anti-CD31 antibody [P2B1]

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-CD31 antibody [P2B1] (AB24590)

IHC image of CD31 staining in a section of frozen normal human placenta performed on a Leica BOND™ system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab24590, 0.1ugml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• ICC

Lab

Immunocytochemistry - Anti-CD31 antibody [P2B1] (AB24590)

ab24590 staining CD31 in HUVEC cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab24590 at 1μg/mL concentration and ab6046 (Rabbit polyclonal to beta Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor^® 488) (ab150117) at 2 μg/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor^® 594) (ab150080) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.

Image was taken with a high-content analysis system (Perkin Elmer, Operetta CLS™).

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-CD31 antibody [P2B1] (AB24590)

Immunohistochemical analysis of frozen human placenta labelling CD31 with ab24590 at a concentration of 0.1µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. ab24590 Anti-FMRP antibody was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)

• IHC

CiteAb

Immunohistochemistry - Anti-CD31 antibody [P2B1] (AB24590)

CD31 immunohistochemistry-immunofluorescence using Anti-CD31 antibody [P2B1] ab24590. Publication image and figure legend from Yuan, L. & Liu, X., 2015, Mol Med Rep, Pubmed 25502723.

Colocalization of the platelet marker CD41 (green) with (A) CD31 (red) or (B) VEGF (red) in human ovarian mucinous cystadenocarcinoma. Tissues (4μm) were incubated with anti-CD41, CD31 and VEFG antibodies prior to staining with fluorescein isothiocyanate and rhodamine-conjugated secondary antibodies. The nuclei were stained using diamidinophenylindole (blue). The samples were analyzed by fluorescence microscopy (magnification, 400x). CD41, platelet glycoprotein IIb;;CD31, platelet endothelial cell adhesion molecule 1; VEGF, vascular endothelial growth factor.

• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-CD31 antibody [P2B1] (AB24590)

CD31 Immunocytochemistry-immunofluorescence using Anti-CD31 antibody [P2B1] ab24590. Publication image and figure legend from Zhan, P., Cui, Y., et al., 2022, Cell Commun Signal, Pubmed 36229856.

COX2 was highly expressed in the laser-injured RPE-choroid complex. A, B Immunofluorescence staining of the choroidal flat mounts A and sections B from mice (7 dpi) with anti-CD31, anti-COX2, and anti-COX1 mAbs; scale bar : 100 μm. C, D The COX2 and COX1 protein at 7 dpi was measured by western blotting using β-actin as an internal control. The representative blots are shown, with quantification (n = 6). E Quantitative analysis of PGE2 levels (n = 6). The level of PGE2 in the peripheral blood of mice (7 dpi) was measured by ELISA after the laser-induced CNV model. *p < < 0.05; **p < < 0.01

• WB

CiteAb

Western blot - Anti-CD31 antibody [P2B1] (AB24590)

CD31 western blotting using Anti-CD31 antibody [P2B1] ab24590. Publication image and figure legend from Li, B., Zhang, T., et al., 2022, Elife, Pubmed 35001873.

Methyltransferase like 3 (METTL3) deficiency induces endothelial activation in atheroprone regions.(A-C) Protein was extracted from the AA and TA of 8-week-old EC-Mettl3KO and Mettl3flox/flox mice. (A) Western blot analysis of the expression of METTL3, vascular adhesion molecule (VCAM-1), CD31, SMA (smooth muscle actin), and GAPDH in tissue lysates of AA and TA intima. AA, aortic arch; TA, thoracic aorta. (B-C) Quantification of protein expression in (A). Data are shown as the mean ± SEM, *p<0.05 (two-way ANOVA with Bonferroni multiple comparison post hoc test). Protein extracts of intima from three mice were pooled as one sample, n = 3. (D) EC-Mettl3KO and Mettl3flox/flox mice underwent partial ligation of the carotid artery for 2 weeks. En face immunofluorescence staining for the expression of VCAM-1 and METTL3 in ECs of the carotid artery of mice. Scale bar, 20 μm. (E) Quantification of the relative fluorescence intensity of VCAM-1 and METTL3. Data are shown as the mean ± SEM, *p<0.05 (two-way ANOVA with Bonferroni multiple comparison post hoc test). n = 6 mice. (F) Male mice underwent partial ligation of the carotid artery. During ligation, carotid arteries were infused with the indicated adeno-associated virus. Immunofluorescence staining of VCAM-1 and vWF in ECs of the RCA and LCA of mice. RCA, right carotid artery; LCA, left carotid artery. Scale bar, 20 μm. (G) Quantification of the relative fluorescence intensity of VCAM-1. Data are shown as the mean ± SEM, *p<0.05 (one-way ANOVA with Bonferroni multiple comparison post hoc test). n = 5 mice.Figure 2'source data 1.Mettl3 deficiency induces endothelial activation in atheroprone regions.Mettl3 deficiency induces endothelial activation in atheroprone regions.Identification of EC-specific methyltransferase like 3 (Mettl3)-deficient mice.(A) Genotyping of mice. PCR indicated that tamoxifen-stimulated Cdh5-Cre recombination in tail preparations of mice as follows : wild-type mice and Mettl3flox/-, Mettl3flox/flox, and EC-Mettl3KO mice. (B) Body weight of 8-week-old EC-Mettl3KO and Mettl3flox/flox mice. Data are shown as the mean ± SEM, NS, not significant (Student's t test). n = 5. (C) Male EC-Mettl3KO mice underwent partial ligation of the carotid artery. During ligation, carotid arteries were infused with the indicated adeno-associated viruses. Immunofluorescence staining for expression of Mettl3 and vWF in ECs of the carotid artery of mice. Scale bar, 20 μm. (D) Quantification of the relative fluorescence intensity of Mettl3. Data are shown as the mean ± SEM, *p<0.05 (Student's t test). n = 5 mice.Figure 2 figure supplement 1 source data 1.Identification of EC-specific Mettl3-deficient mice.Identification of EC-specific Mettl3-deficient mice.

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