Anti-Carbonic Anhydrase 9/CA9 antibody [EPR4151(2)] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Carbonic Anhydrase 9/CA9 antibody [EPR4151(2)] - BSA and Azide free (AB180539)

This data was developed using ab180539, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human stomach tissue sections labeling Carbonic Anhydrase 9/CA9 with Purified ab180539 at 1 : 150 dilution (0.89 μg/ml). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• WB

Lab

Western blot - Anti-Carbonic Anhydrase 9/CA9 antibody [EPR4151(2)] - BSA and Azide free (AB180539)

This data was developed using the same antibody clone in a different buffer formulation (ab108351).

Western blot : Anti-CA9 antibody [EPR4151(2)] (ab108351) staining at 1/200 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab108351 was shown to bind specifically to CA9. A band was observed at 50 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in CA9 knockout cell line. To generate this image, wild-type and CA9 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Carbonic Anhydrase 9/CA9 antibody [EPR4151(2)] (<a href='/en-us/products/primary-antibodies/carbonic-anhydrase-9-ca9-antibody-epr41512-ab108351'>ab108351</a>) at 1/200 dilution

Lane 1:

Wild-type HCT 116 cell lysate at 20 µg

Lane 2:

CA9 knockout HCT 116 cell lysate at 20 µg

Lane 3:

MCF7 cell lysate at 20 µg

Lane 4:

HT-29 cell lysate at 20 µg

Lane 5:

Jurkat cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 50 kDa

false

• WB

Unknown

Western blot - Anti-Carbonic Anhydrase 9/CA9 antibody [EPR4151(2)] - BSA and Azide free (AB180539)

This data was developed using ab108351, the same antibody clone in a different buffer formulation.

The molecular weight observed is consistent with what has been described in the literature (PMID : 12966427, 30734594 and 15951534).

Blocking Buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-Carbonic Anhydrase 9/CA9 antibody [EPR4151(2)] (<a href='/en-us/products/primary-antibodies/carbonic-anhydrase-9-ca9-antibody-epr41512-ab108351'>ab108351</a>) at 1/10000 dilution

All lanes:

Human stomach lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 49 kDa

Observed band size: 54 kDa,58 kDa

false

• WB

Unknown

Western blot - Anti-Carbonic Anhydrase 9/CA9 antibody [EPR4151(2)] - BSA and Azide free (AB180539)

This data was developed using ab108351, the same antibody clone in a different buffer formulation.

The molecular weight observed is consistent with what has been described in the literature (PMID : 12966427, 30734594 and 15951534).

Blocking Buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-Carbonic Anhydrase 9/CA9 antibody [EPR4151(2)] (<a href='/en-us/products/primary-antibodies/carbonic-anhydrase-9-ca9-antibody-epr41512-ab108351'>ab108351</a>) at 1/10000 dilution

All lanes:

HT-29 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 49 kDa

Observed band size: 54 kDa,58 kDa

false