Anti-CD31 antibody

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody (AB124432)

IHC image of CD31 staining in mouse placenta formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer pH6 for 20 mins. The section was then incubated with ab124432, 1μg/ml, for 15 mins at room temperature. A goat anti-rabbit biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC 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.

• IHC-P

PubMed

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD31 antibody (AB124432)

Formalin-fixed, paraffin-embedded ischemic and normal mouse hindlimb tissue stained for CD31 using ab124432 at 1/1000 dilution. The Rabbit specific HRP/DAB detection kit (ab64261) was used as the secondary reagent. CD31 staining reveals the formation of larger intramuscular blood vessels in limbs receiving (h) both LPA and ASC, while (e) normal tissue and defects receiving (f) LPA and (g) ASC alone have fewer and smaller vessels.

Binder BY et al. Lysophosphatidic acid enhances stromal cell-directed angiogenesis. PLoS One 8:e82134 (2013). Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

• WB

Lab

Western blot - Anti-CD31 antibody (AB124432)

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab124432 overnight at 4°C. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) at a 1 : 10000 dilution for 1hr at room temperature and then imaged.

All lanes:

Western blot - Anti-CD31 antibody (ab124432) at 1 µg/mL

Lane 1:

TH2 whole cell lysate at 20 µg

Lane 2:

Mouse spleen at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 82 kDa

Observed band size: 120-140 kDa

false

• WB

Unknown

Western blot - Anti-CD31 antibody (AB124432)

Abcam recommends using milk as the blocking agent - 3%. CD31 contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted. This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab124432 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

All lanes:

Western blot - Anti-CD31 antibody (ab124432) at 1 µg/mL

Lane 1:

Heart (Mouse) Tissue Lysate at 25 µg

Lane 2:

Spleen (Mouse) Tissue Lysate at 25 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/10000 dilution

Predicted band size: 82 kDa

Observed band size: 130 kDa,90 kDa

true

Exposure time: 20min

• WB

CiteAb

Western blot - Anti-CD31 antibody (AB124432)

CD31 western blot using anti-CD31 antibody ab124432. Publication image and figure legend from Wang, J., Niu, N., et al., 2019, Sci Rep, PubMed 30728372.

ab124432 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab124432 please see the product overview.

Identification of cultured MLECs by western blot and confocal microscopy. (A,B) RNA-sequencing data mined from published literature19 shows expression abundance of three EC marker genes, VE-cadherin (also known as CDH5), CD31 (also known as PECAM1), and vWF. Data shown are normalized counts from two different donors of HUVECs. (C) CD31 expression of EC fraction (CD31+; ICAM2+) and non-EC (CD31−; ICAM2−) fraction after second bead sorting, n = 3 different mice. (D) VE-cadherin (red) and vWF (green) staining of cultured MLEC by confocal microscopy, n = 4, scale bar = 60 um. (E), DiI-oxLDL uptake (red) by cultured MLECs, n = 5, scale bar = 60 um.

false