重组Anti-Carbonic anhydrase 2/CA2抗体[EPR5195] (ab124687)

Lane 1: Wild-type HAP1 cell lysate (40 µg)
Lane 2: Carbonic anhydrase 2/CA2 knockout HAP1 cell lysate (40 µg)
Lane 3: A431 cell lysate (40 µg)
Lane 4: HEK293 cell lysate (40 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab124687 observed at 32 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab124687 was shown to specifically react with Carbonic anhydrase 2/CA2 when Carbonic anhydrase 2/CA2 knockout samples were used. Wild-type and Carbonic anhydrase 2/CA2 knockout samples were subjected to SDS-PAGE. Ab124687 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10,000 dilution respectively and incubated overnight at 4C. Blots were developed with IRDye^® 800CW Goat anti-Rabbit IgG (H + L) and IRDye® 680 Goat anti-Mouse IgG (H + L) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue sections labeling Carbonic anhydrase 2/CA2 with purified ab124687 at 1/1600 (0.063 µg/ml). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Lanes 1-4: Merged signal (red and green). Green - ab124687 observed at 29 kDa. Red - loading control ab8245 observed at 36 kDa. 

 ab124687 Anti-Carbonic anhydrase 2/CA2 antibody [EPR5195] was shown to specifically react with Carbonic anhydrase 2/CA2 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab265072 (knockout cell lysate ab257084) was used.  Wild-type and Carbonic anhydrase 2/CA2 knockout samples were subjected to SDS-PAGE.  ab124687 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kindey tissue sections labeling Carbonic anhydrase 2/CA2 with purified ab124687 at 1/1600 (0.063 µg/ml). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labeling Carbonic anhydrase 2/CA2 with Purified ab124687 at 1/1600 (0.063 µg/ml). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human clear cell carcinoma tissue sections labeling Carbonic anhydrase 2/CA2 with purified ab124687 at 1/1600 (0.063 µg/ml). Heat mediated antigen retrieval was performed Bond™ Epitope Retrieval Solution 2 (pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.