Anti-S100A4 antibody (ab41532)

Western blot - S100A4 antibody (ab41532)

Anti-S100A4 antibody (ab41532) at 1 µg/ml + S100A4 - Recombinant Protein at 0.1 µg

Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Performed under reducing conditions.

Predicted band size : 12 kDa
Observed band size : 37 kDa (why is the actual band size different from the predicted?)


The S100A4 tagged recombinant protein has a molecular weight of 36.8 kDa.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - S100A4 antibody (ab41532)

ab41532 staining S100A4 in human heart tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent fixation in formaldehyde, heat mediated antigen retrieval in Citrate buffer pH 6.0 and blocking (5 minutes/peroxidase block then 10 minutes/protein block) for 15 minutes at 20°C. The primary antibody was diluted, 1/2000 and incubated with sample for 45 minutes at 20°C. A HRP conjugated goat polyclonal to rabbit IgG was used undiluted as secondary.

This image is courtesy of an Abreview submitted by Antibody Solutions Ltd.

Immunocytochemistry/ Immunofluorescence - S100A4 antibody (ab41532)

ICC/IF image of ab41532 stained HepG2 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 (ab41532, 5µg/ml) overnight at +4ºC. 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. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa, Hek293 and MCF7 cells at 5µg/ml, and in 100% methanol fixed (5 min) HeLa, Hek293, HepG2 and MCF7 cells at 5µg/ml.

Western blot - Anti-S100A4 antibody (ab41532)

Anti-S100A4 antibody (ab41532) at 1/250 dilution + S100A4 protein (Human) (ab83650) at 0.01 µg

Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (ab97080) at 1/5000 dilution
developed using the ECL technique

Performed under reducing conditions.

Exposure time : 20 minutes