Immunohistochemical analysis of paraffin-embedded Human stomach tissue labeling Carbonic Anhydrase 9/CA9 with ab243660 at 1/2000 dilution (0.262 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Membranous staining on human stomach (PMID: 15951534) is observed. The section was incubated with ab236628 for 10 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 solution2) for 20 mins.

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).

Immunohistochemical analysis of 4% PFA fixed 0.2% Triton X-100 permeabilized frozen Rat stomach tissue labeling Carbonic Anhydrase 9/CA9 with ab243660 at 1/100 dilution followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution. The nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody was ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution.

All lanes : Anti-Carbonic Anhydrase 9/CA9 antibody [EPR23055-5] (ab243660) at 1/1000 dilution

Lane 1 : Mouse colon tissue lysate
Lane 2 : Mouse brain tissue lysate
Lane 3 : Mouse spleen tissue lysate
Lane 4 : Rat colon tissue lysate
Lane 5 : Rat brain tissue lysate
Lane 6 : Rat spleen tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 49 kDa


Exposure time: 15 seconds

Blocking/Diluting buffer and concentration: 5% NFDM/TBST

Exposure time: 15 seconds.

Immunohistochemical analysis of paraffin-embedded Rat stomach tissue labeling Carbonic Anhydrase 9/CA9 with ab243660 at 1/2000 dilution (0.262 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Membranous staining on rat stomach is observed. The section was incubated with ab236628 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 solution2) for 20 mins.

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).

Immunohistochemical analysis of paraffin-embedded Mouse stomach tissue labeling Carbonic Anhydrase 9/CA9 with ab243660 at 1/2000 dilution (0.262 μg/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Membranous staining on mouse stomach is observed. The section was incubated with ab236628 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 solution2) for 20 mins.

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).

Immunohistochemical analysis of 4% PFA fixed 0.2% Triton X-100 permeabilized frozen Mouse stomach tissue labeling Carbonic Anhydrase 9/CA9 with ab243660 at 1/100 dilution followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 (2 μg/ml) dilution. The nuclear counterstain was DAPI (Blue). Positive staining on mouse stomach tissue is observed.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody was ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1:1000 (2 μg/ml) dilution.

All lanes : Anti-Carbonic Anhydrase 9/CA9 antibody [EPR23055-5] (ab243660) at 1/1000 dilution

Lane 1 : L-929 ( mouse connective tissue fibroblast) whole cell lysate
Lane 2 : A20 ( mouse reticulum sarcoma b lymphocyte) whole cell lysate
Lane 3 : 4T1 ( mouse mammary gland carcinoma epithelial cell) whole cell lysate
Lane 4 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 49 kDa


Exposure time: 1 second

Blocking/Diluting buffer and concentration: 5% NFDM/TBST

Exposure time: 1 second.

Carbonic Anhydrase 9/CA9 was immunoprecipitated from 0.35 mg 4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate with ab243660 at 1/30 dilution. Western blot was performed on the immunoprecipitate using 243660 at 1/1000 dilution (0.52 μg/ml). VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: 4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate 10 μg

Lane 2: ab243660 IP in 4T1 whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab243660 in 4T1 whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 48