Anti-CD31抗体(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.

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.

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.