Anti-HSD17B1 antibody [EP1682Y] (ab51045) at 1/100000 dilution + Human placenta lysate at 10 µg

Secondary
Goat anti-Rabbit HRP labeled. at 1/2000 dilution

Predicted band size: 35 kDa
Observed band size: 35 kDa

Ab51045 (1/100 dilution) staining human HSD17B1 in paraffin embedded human placenta tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Purified ab51045 at 1/50 dilution (2µg) immunoprecipitating HSD17B1 in Human placenta lysate.
Lane 1 (input): Human placenta lysate 10µg
Lane 2 (+): ab51045 + Human placenta lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab51045 in Human placenta lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 34 kDa

Overlay histogram showing JEG3 cells stained with ab51045 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab51045, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x10⁶ cells ) used under the same conditions. Acquisition of >5,000 events was performed.