Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: IP3 receptor knockout HAP1 cell lysate (20 µg) 
Lane 3: Human brain tissue lysate (20 µg)
Lane 4: SH-SY5Y cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab108517 observed at 270 kDa. Red - loading control, ab18058, observed at 124 kDa.

ab108517 was shown to specifically recognize IP3 receptor in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when IP3 receptor knockout samples were examined. Wild-type and IP3 receptor knockout samples were subjected to SDS-PAGE. ab108517 and ab18058 (loading control to Vinculin) were diluted 1/1000 and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108517).

Immunohistochemical analysis of paraffin-embedded Human cerebellum  tissue labeling IP3 with ab108517 at 1/1000 dilution (0.139 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human cerebellum. The section was incubated with ab108517 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108517).

Immunohistochemical analysis of paraffin-embedded Mouse cerebellum tissue labeling IP3 with ab108517 at 1/1000 dilution (0.139 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse cerebellum. The section was incubated with ab108517 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20mins.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108517).

ab108517 (purified) at 1:500 dilution (1.124 µg/ml) immunoprecipitating IP3 receptor in Mouse brain lysate.
Lane 1 (input): Mouse brain lysate 10µg
Lane 2 (+): ab108517 & Mouse brain lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32061 in HeLa whole cell lysate
For western blotting, VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM /TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108517).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108517).