Postconfluent HUVECs in 96-well plates were treated for 30 min with comfrey extracts and then stimulated with IL-1β (5 ng/ml) in the same medium. After 2 h, cells were fixed with 4% paraformaldehyde for 15 min following a blocking with 2.5% bovine serum albumin (BSA) in Tris-Buffered Saline-1% Tween (TBS-T) for 1 h. Afterward, cells were incubated overnight at 4°C with mouse-anti-E-selectin antibody (R&D Systems) diluted 1:500 in 1% BSA/TBS-T. Following washing with TBS-T, goat-anti-mouse horseradish-peroxidase (HRP) antibody (Sigma) diluted 1:10,000 in 1% BSA/TBS-T was added for 1 h. HRP activity was assessed using tetramethylbenzidine (Sigma) as substrate. The reaction was stopped adding 2 M H2SO4 and absorbance measured at OD450nm. E-selectin levels were normalized to crystal violet staining.
Mouse anti e selectin antibody
Manufactured by R&D Systems
The Mouse-anti-E-selectin antibody is a laboratory reagent used in various research applications. E-selectin is a cell adhesion molecule expressed on the surface of endothelial cells and plays a role in the recruitment of leukocytes to sites of inflammation. This antibody can be used to detect and quantify E-selectin expression in a range of in vitro and in vivo experimental models.
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Lab products found in correlation
2 protocols using mouse anti e selectin antibody
1
Comfrey Extract Modulation of E-selectin
2
Evaluating Comfrey Extract's Inhibition of E-Selectin
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Post-confluent human umbilical vein endothelial cells (HUVEC) in 96-well plates were treated for 30 min with comfrey extract samples and then stimulated with IL-1β (5 ng/mL) in the same medium. After 2 h, cells were fixed with 4% paraformaldehyde for 15 min after blocking with 2.5% bovine serum albumin (BSA) in Tris-buffered saline-1% tween (TBS-T) for 1 h. The cells were then incubated overnight at 4 • C with mouse anti-E-selectin antibody (R&D Systems) diluted 1:500 in 1% BSA/TBS-T. After washing with TBS-T for 1 h, goat antihorseradish peroxidase (HRP) antibody (Sigma) diluted 1:10,000 in 1% BSA/TBS-T was added. HRP activity was assessed using tetramethylbenzidine (Sigma) as substrate. The addition of 2M H 2 SO 4 stopped the reaction, and the optical density OD 450 was measured. E-selectin levels were normalized by crystal violet staining.
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