Recombinant Anti-beta Catenin antibody [E247] - BSA and Azide free (ab196204)

This data was developed using the same antibody clone in a different buffer formulation (ab32572).

Lanes 1-4: Merged signal (red and green). Green - ab32572 observed at 95 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab32572 was shown to react with beta Catenin in wild-type HeLa. Loss of signal was observed when knockout sample ab263756 was used. Wild-type and beta Catenin knockout samples were subjected to SDS-PAGE. ab32572 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 5000 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.

Different expression level of beta Catenin in HCTs (hepatocellular carcinoma tissues) and PLTs (para-cancerous liver tissues).

The HCTs, PLTs were paraffin-embedded and cut into sections with 5 μm-thickness for hematoxylin-eosin and immunohistochemistry (IHC) analysis. ab32572 was used at a dilution of 1:400. The second antibody was a biotinylated IgG to incubate 40 minutes at 37°C. Finally, the tissue slices were visualized by the 3, 3-diaminobenzidine solution and counterstained with hematoxylin. Substitution of the primary antibody with phosphate-buffered saline was served as a control for IHC.

The beta Catenin with negative, weak, moderate and strong staining activity was respectively detected in HCTs (E-H) and PLTs (M-P). Section E shown above, for full image please see original paper.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32572).

ab32572 staining in CTNNB1 (beta Catenin) wild-type HAP1 cells (top panel) and in CTNNB1 (β-catenin) knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab32572 at 1/250 dilution and ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody at 2 μg/ml (shown in green). Nuclear DNA was labeled in blue with DAPI.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32572).

ab32572 staining beta Catenin in the bEnd.5 murine cell line by ICC/IF (Immunocytochemistry/immunofluorescence).

Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton in PBS and blocked with 10% serum for 30 minutes at 22°C. Samples were incubated with primary antibody (1/300) for 16 hours at 4°C. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32572).

ab32572 staining beta Catenin in SW480 (Human colorectal adenocarcinoma cell line) cells treated with BIO (ab120891), by ICC/IF. Increase of beta Catenin expression correlates with increased concentration of BIO, as described in literature.
The cells were incubated at 37°C for 48h in media containing different concentrations of ab120891 (BIO) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab32572 (1/200) dilution was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A Goat Anti-Rabbit IgG H&L (DyLight^® 488) preadsorbed (ab96899) secondary antibody at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32572).

ab32572 staining beta Catenin in dog colon tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).

Tissue was fixed with formaldehyde and blocked with 10% serum for 30 minutes at 25°C; antigen retrieval was by heat mediation in 10 mM citrate buffer, pH6. Samples were incubated with primary antibody (1/250 in PBS with 1x casein) for 90 minutes at 25°C. A biotin-conjugated Goat anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32572).

ab32572 at 1/200 staining mouse small intestine tissue sections by IHC-P.

The tissue was formaldehyde fixed and a heat mediated antigen retrieval step was performed before incubation with the primary antibody. An HRP conjugated goat anti-rabbit antibody was used as the secondary.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32572).

ab32572 staining human renal carcinoma tissue sections by IHC-P. 

Sections were formaldehyde fixed and subjected to heat mediated antigen retrieval in citrate buffer (pH 6) prior to blocking with 1% milk for 45 minutes at 22°C.  The primary antibody was diluted 1/200 and incubated with the sample for 1 hour at 22°C.  An HRP conjugated goat anti-rabbit antibody, diluted 1/400, was used as the secondary.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32572).

ab32572 staining beta Catenin in mouse liver tissue section by Immunohistochemistry (Frozen sections).

Tissue samples were fixed with formaldehyde and blocked with 5% serum at 40C for 30 minutes. The sample was incubated with primary antibody (1/200) in dilution buffer containing PBS and 3% goat serum at 40C for 9 hours. An Alexa Fluor^®488-conjugated Goat polyclonal to rabbit IgG was used as secondary antibody at 1/200 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32572).

ab32572 showing positive staining in human cervical carcinoma tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32572).

ab32572 showing positive staining in human breast carcinoma tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32572).

ab32572 showing positive staining in human lung adenocarcinoma tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32572).

ab32572 showing positive staining in human papillary carcinoma of thyroid gland tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32572).

ab32572 showing positive staining in human kidney carcinoma tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32572).

ab32572 staining beta Catenin in SK-N-SH (Human neuroblastoma cell line) cells treated with olanzapine (ab120736), by ICC/IF.

Increase in expression of beta Catenin correlates with increased concentration of olanzapine, as described in literature.
The cells were incubated at 37°C for 24h in media containing different concentrations of ab120736 (olanzapine) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab32572 (1/200 dilution) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A Goat Anti-Rabbit IgG H&L (DyLight^® 488) preadsorbed (ab96899) secondary antibody at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32572).