Anti-BCRP/ABCG2 antibody [BXP-53] (ab24115)

Western blot - BCRP/ABCG2 antibody [BXP-53] (ab24115)

All lanes : Anti-BCRP/ABCG2 antibody [BXP-53] (ab24115) at 1 µg/ml

Lane 1 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate
Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate

Lysates/proteins at 10 µg per lane.

Secondary
Peroxidase Conjugated AffiniPure Rabbit Anti-Rat IgG (H+L) at 1/10000 dilution
developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 72 kDa
Observed band size : 75 kDa (why is the actual band size different from the predicted?)
Additional bands at : 37 kDa. We are unsure as to the identity of these extra bands.

Exposure time : 20 minutes

BCRP/ABCG2 contains a potential glycosylation site (SwissProt) which may explain its migration at a higher molecular weight than predicted.

Flow Cytometry-BCRP/ABCG2 antibody [BXP-53](ab24115)

Overlay histogram showing HEK293 cells stained with ab24115 (red line). The cells were fixed with 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 followed by the antibody (ab24115, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rat IgG (Fc) (ab96971) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rat IgG2a [aRTK2758] (ab18450, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a significantly decreased signal in HEK293 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

Immunocytochemistry/ Immunofluorescence - BCRP/ABCG2 antibody [BXP-53] (ab24115)

ab24115 staining BCRP/ABCG2 in murine mesenchymal stem cells by Immunocytochemistry/ Immunofluorescence. The cells were fixed in formaldehyde, permeabilised in 0.1% Triton X and then blocked using 1% serum for 1 hour at 25ºC. Samples were then incubated with primary antibody at 1/25 for 15 hours at 25ºC. The secondary antibody used was a donkey anti-rat IgG conjugated to an Alexa Fluor® used at a 1/100 dilution.

Image courtesy of an anonymous Abreview.