Anti-LGALS3BP antibody (ab67353)

All lanes : Anti-LGALS3BP antibody (ab67353) at 1/500 dilution

Lane 1 : LGALS3BP transfected 293T cell lysate
Lane 2 : Non transfected 293T cell lysate

Lysates/proteins at 25 µg per lane.

Secondary
All lanes : Goat Anti-Mouse IgG (H&L)-HRP Conjugate at 1/2500 dilution

Predicted band size: 65 kDa
Observed band size: 80 kDa why is the actual band size different from the predicted?

IHC image of ab67353 staining in Breast Cancer formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab67353, 5µg/ml, 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/IF image of ab67353 stained HeLa cells. The cells were 4% formaldehyde (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab67353, 1µg/ml) overnight at +4°C. The secondary antibody (green) was for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.