Anti-CD62E 抗体 [EPR25616-62] (BSA and Azide free) (ab300558)

Anti-CD62E antibody [EPR25616-62] (ab300557) at 1/1000 dilution + Human CD62E active protein, 10 ng

Secondary
Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Developed using the ECL technique.

Observed band size: 115 kDa why is the actual band size different from the predicted?


Exposure time: 180 seconds

This data was developed using ab300557, the same antibody clone in a different buffer formulation. 

Blocking and diluting buffer and concentration: 5% NFDM/TBST

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes : Anti-CD62E antibody [EPR25616-62] (ab300557) at 1/1000 dilution

Lane 1 : Untreated HUVEC (human umbilical vein endothelial cell), whole cell lysate
Lane 2 : HUVEC treated with 20ng/ml IL-lbeta for 4h, whole cell lysate

Lysates/proteins at 50 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Developed using the ECL technique.

Observed band size: 115 kDa why is the actual band size different from the predicted?


Exposure time: 81 seconds

This data was developed using 300557, the same antibody clone in a different buffer formulation. 

Blocking and diluting buffer and concentration: 5% NFDM/TBST

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

The molecular weight observed is consistent with what has been described in the literature (PMID: 16715142; PMID: 7589110). 

This data was developed using ab300557, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HUVEC (human umbilical vein endothelial cells) labeling CD62E with ab300557 at 1/50 dilution (9.36 μg/ml), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (2 μg/ml) (Green). Confocal image showing increased cytoplasmic and membrane staining in HUVEC cells treated with interleukin 1-lbeta (20 ng/ml) for 4 h (PMID: 16715142). 

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilutioin (2.5 μg/mL) (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only control: PBS was used instead of primary antibody, followed by a preadsorbed secondary antibody ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 (2 μg/ml). HUVEC cells treated and untreated with interleukin 1-beta (20 ng/ml) for 4 h were tested.  

This data was developed using ab300557, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of HUVEC (human umbilical vein endothelial cell) treated with 20 ng/ml interleukin 1 beta for 4 hours (Right) / Untreated control (Left) cells labelling CD62E with ab300557 at 1/500 dilution (0.1 μg). Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.