All lanes : Anti-P2Y6 antibody [EPR3816] (ab92504) at 1/20000 dilution

Lane 1 : A549 cell lysate
Lane 2 : HACAT cell lysate
Lane 3 : HepG2 cell lysate
Lane 4 : Human spleen lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP-labeled goat anti-rabbit at 1/2000 dilution

Predicted band size: 36 kDa

This data was developed using ab92504, the same antibody clone in a different buffer formulation.

This data was developed using the same antibody clone in a different buffer formulation (ab92504).
Immunocytochemistry analysis of A549 (human lung carcinoma epithelial cell) labeling P2Y6 with purified ab92504 at 1/100 dilution. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/1000 (2 µg/ml) was used as the secondary antibody. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.20 µg/ml) was used as counterstain. Nuclei were stained blue with DAPI.
Negative control: PBS instead of the primary antibody.

This data was developed using ab92504, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Human kidney tissue (left-hand image) and Human liver tissue (right-hand image) using ab92504 at a dilution of 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

This data was developed using ab92504, the same antibody clone in a different buffer formulation.
Purified ab92504 at 1/50 dilution (2µg) immunoprecipitating P2Y6 in HaCaT whole cell lysate.
Lane 1 (input): HaCaT (Human skin keratinocyte) whole cell lysate 10µg
Lane 2 (+): ab92504 + HaCaT whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab92504 in HaCaT whole cell 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: 42 kDa