Anti-Galectin 1 antibody (ab25138) at 1 µg/ml + Human placenta tissue lysate - total protein (ab29745) at 10 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 14 kDa
Observed band size: 14 kDa
Additional bands at: 32 kDa, 45 kDa, 73 kDa. We are unsure as to the identity of these extra bands.


Exposure time: 30 seconds

ICC/IF image of ab25138 stained Hek293 cells. The cells were 4% PFA fixed (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 (ab25138, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution 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.

IHC image of ab25138 staining in human normal kidney 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 ab25138, 1µg/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.

ab25138 staining Galectin-1 in Human cervical cancer tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Samples were incubated with primary antibody (1/1000 in 1% BSA in PBS) overnight at room temperature. An Alexa Fluor A647-conjugated Donkey anti-rabbit polyclonal (1/200) was used as the secondary antibody.

Representative images from double stainings of Galectin-1 and CD163. Arrows indicate examples of double positive cells.

ab25138 staining Galectin-1 in Human cervical cancer tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Samples were incubated with primary antibody (1/1000 in 1% BSA in PBS) overnight at room temperature. An Alexa Fluor A647-conjugated Donkey anti-rabbit polyclonal (1/200) was used as the secondary antibody.

Representative images from double stainings of Galectin-1 and CD3.