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AB196204

Anti-beta Catenin antibody [E247] - BSA and Azide free

  • BOND RX™ Validated
  • RabMAb
  • Recombinant
  • KO Validated
  • Advanced Validation
  • What is this?

5

(1 Review)

|

(34 Publications)

Anti-beta Catenin antibody [E247] - BSA and Azide free (ab196204) is a rabbit recombinant monoclonal antibody in a PBS only buffer for easy conjugation detecting beta Catenin in Western Blot, IP, IHC-P, ICC/IF, ChIP. Suitable for Human, Mouse, Rat.

- KO validated for confirmed specificity
- BSA, sodium azide, and glycerol-free for easy conjugation
- Biophysical QC for unrivalled batch-batch consistency
- Over 30 publications

View Alternative Names

CTNNB, OK/SW-cl.35, PRO2286, CTNNB1, Catenin beta-1, Beta-catenin

28 Images
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)
  • IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)

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).

Immunocytochemistry/ Immunofluorescence - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)
  • ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)

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).

Immunocytochemistry/ Immunofluorescence - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)
  • ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)

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).

Immunocytochemistry/ Immunofluorescence - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)
  • ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)

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).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)
  • IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)

Immunohistochemical analysis of formalin fixed paraffin embedded human tonsil labelling beta catenin with ab196204 at a concentration of 1µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32 min with ULTRA cell conditioning solution (CC1 pH 8.5). ab196204 anti beta catenin antibody was incubated at 37°C for 16min. Sections were counterstained is with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)
  • IHC-P

PubMed

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)

Different expression level of beta Catenin in Human 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).

Image from Jin et al PLoS One. 2015 Aug 7;10(8):e0133770. doi: 10.1371/journal.pone.0133770. eCollection 2015. Fig 2.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)
  • IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)

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).

Immunocytochemistry/ Immunofluorescence - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)
  • ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)

Clone E247 (ab196204) has been successfully conjugated by Abcam. This image was generated using Anti-beta Catenin antibody [E247] (Alexa Fluor® 488). Please refer to ab194118 for protocol details.

ab194118 staining β-catenin in wild-type HAP1 cells (top panel) and 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 ab194118 at 1/500 dilution (shown in green) and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.

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

Immunocytochemistry/ Immunofluorescence - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)
  • ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)

Clone E247 (ab196204) has been successfully conjugated by Abcam. This image was generated using Anti-beta Catenin antibody [E247] (Alexa Fluor® 647). Please refer to ab194119 for protocol details.

ab194119 staining β-catenin in wild-type HAP1 cells (top panel) and 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 ab194119 at 1/500 dilution (shown in red) and ab195887 at 1/250 dilution (shown in green) overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.

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

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)
  • IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)

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).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)
  • IHC-P

AbReview8673****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)

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).

This image is courtesy of an anonymous Abreview.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)
  • IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)

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).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)
  • IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)

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).

ChIP - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)
  • ChIP

Lab

ChIP - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)

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

Chromatin was prepared from HCT 116 cells according to the Abcam Dual-X-ChIP protocol*. Cells were fixed with 1.5 mM EGS for 30 mins and then formaldehyde for 10 min.

The ChIP was performed with 25 μg of chromatin, 5 μg of ab32572 (red), or 5 μg of rabbit normal IgG ab172730 (gray) and 20 μl of Protein A/G sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci)

Primers and probes are from paper PMID : 28625518

*http : //www.abcam.com/resources?keywords=X%20ChIP%20protocol

Multiplex immunohistochemistry - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)
  • mIHC

Lab

Multiplex immunohistochemistry - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)

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

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded rat liver tissue staining ALDH1L1 with ab307696 at a 1/5000 dilution, ab302928 anti-HIRA/HIR used at 1/100 dilution and ab32572 anti-beta Catenin used at a 1/250 dilution. Opal Polymer HRP Ms + Rb was used as a secondary antibody.

Panel A : merged staining of anti-ALDH1L1 (green; Opal520), anti-HIRA/HIR (magenta; Opal690) and anti-beta Catenin (grey; Opal570) on rat liver.
Panel B : anti-ALDH1L1 staining cytoplasm of hepatocytes in rat liver.
Panel C : anti-HIRA/HIR staining nucleus of hepatocytes in rat liver.
Panel D : anti-beta Catenin staining membrane of hepatocytes in rat liver.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab307696, ab302928 and ab32572 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Immunocytochemistry/ Immunofluorescence - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)
  • ICC/IF

AbReview51865****

Immunocytochemistry/ Immunofluorescence - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)

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).

This image is courtesy of an anonymous Abreview.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)
  • IHC-P

AbReview8674****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)

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).

This image is courtesy of an anonymous Abreview.

Multiplex immunohistochemistry - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)
  • mIHC

Lab

Multiplex immunohistochemistry - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)

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

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse liver tissue staining ALDH1L1 with ab307696 at a 1/5000 dilution, ab302928 anti-HIRA/HIR used at 1/100 dilution and ab32572 anti-beta Catenin used at a 1/250 dilution. Opal Polymer HRP Ms + Rb was used as a secondary antibody.

Panel A : merged staining of anti-ALDH1L1 (green; Opal520), anti-HIRA/HIR (magenta; Opal690) and anti-beta Catenin (grey; Opal570) on mouse liver.
Panel B : anti-ALDH1L1 staining cytoplasm of hepatocytes in mouse liver.
Panel C : anti-HIRA/HIR staining nucleus of hepatocytes in mouse liver.
Panel D : anti-beta Catenin staining membrane of hepatocytes in mouse liver.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab307696, ab302928 and ab32572 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Western blot - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)
  • WB

Supplier Data

Western blot - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)

False colour image of Western blot : Anti-beta Catenin antibody [E247] - ChIP Grade staining at 1/5000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32572 was shown to bind specifically to beta Catenin. A band was observed at 85/90 kDa in wild-type MCF7 cell lysates with no signal observed at this size in CTNNB1 knockout cell line ab286762. To generate this image, wild-type and CTNNB1 knockout MCF7 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. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32572).

All lanes:

Western blot - Anti-beta Catenin antibody [E247] - ChIP Grade (<a href='/en-us/products/primary-antibodies/beta-catenin-antibody-e247-chip-grade-ab32572'>ab32572</a>) at 1/5000 dilution

Lane 1:

Wild-type MCF7 cell lysate at 20 µg

Lane 2:

CTNNB1 knockout MCF7 cell lysate, at 20 µg

Lane 3:

HeLa 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

Predicted band size: 85 kDa

Observed band size: 85/90 kDa

false

Western blot - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)
  • WB

Lab

Western blot - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)

This data was developed using ab32572, the same antibody clone in a different buffer formulation. Different batches of ab32572 were tested on A431 (Human epidermoid carcinoma epithelial cell) lysate at 2.0 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 92 kDa.

All lanes:

Western blot - Anti-beta Catenin antibody [E247] - ChIP Grade (<a href='/en-us/products/primary-antibodies/beta-catenin-antibody-e247-chip-grade-ab32572'>ab32572</a>)

Predicted band size: 85 kDa

false

Western blot - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)
  • WB

Lab

Western blot - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)

Lanes 1 - 4 : Merged signal (red and green). Green - ab32572 observed at 90 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab32572 was shown to specifically react with CTNNB1 (β-catenin) in wild type HAP1 cells. No band was observed when CTNNB1 (β-catenin) knockout samples were used. Wild-type and CTNNB1 (β-catenin) knockout samples were subjected to SDS-PAGE. ab32572 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at a 1/5000 dilution and 1/10000 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 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-beta Catenin antibody [E247] - BSA and Azide free (ab196204) at 1/5000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

CTNNB1 (β-catenin) knockout HAP1 whole cell lysate at 20 µg

Lane 3:

HeLa whole cell lysate at 20 µg

Lane 4:

A431 whole cell lysate at 20 µg

Predicted band size: 85 kDa

false

Western blot - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)
  • WB

Lab

Western blot - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)

False colour image of Western blot : Anti-beta Catenin antibody [E247] - ChIP Grade staining at 1/5000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32572 was shown to bind specifically to beta Catenin. A band was observed at 85 kDa in wild-type HepG2 cell lysates with no signal observed at this size in CTNNB1 knockout cell line (ab277911). To generate this image, wild-type and CTNNB1 knockout HepG2 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-beta Catenin antibody [E247] - BSA and Azide free (ab196204) at 1/5000 dilution

Lane 1:

Wild-type HepG2 cell lysate at 20 µg

Lane 2:

CTNNB1 knockout HepG2 cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

A431 cell lysateA431 cell lysateA431 cell lysateA431 cell lysateA431 cell lysate at 20 µg

Predicted band size: 85 kDa

Observed band size: 85 kDa

false

Western blot - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)
  • WB

Lab

Western blot - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)

False colour image of Western blot : Anti-beta Catenin antibody [E247] - ChIP Grade staining at 1/5000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32572 was shown to bind specifically to beta Catenin. A band was observed at 95 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in CTNNB1 CRISPR-Cas9 edited cell line ab273712 (CRISPR-Cas9 edited cell lysate ab275247). The band observed in the CRISPR-Cas9 edited lysate lane below 95 kDa is likely to represent a truncated form of beta Catenin. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and CTNNB1 CRISPR-Cas9 edited 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 5 % 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-beta Catenin antibody [E247] - ChIP Grade (<a href='/en-us/products/primary-antibodies/beta-catenin-antibody-e247-chip-grade-ab32572'>ab32572</a>) at 1/5000 dilution

Lane 1:

Wild-type HCT 116 cell lysate at 20 µg

Lane 2:

CTNNB1 CRISPR-Cas9 edited HCT 116 cell lysate at 20 µg

Lane 2:

Western blot - Human CTNNB1 knockout HCT116 cell line (<a href='/en-us/products/cell-lines/human-ctnnb1-knockout-hct116-cell-line-ab273712'>ab273712</a>)

Predicted band size: 85 kDa

Observed band size: 95 kDa

false

Western blot - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)
  • WB

Lab

Western blot - 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- 2 : Merged signal (red and green). Green - ab32572 observed at 86 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab32572 was shown to react with beta Catenin in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255352 (knockout cell lysate ab263756) was used. Wild-type HeLa and CTNNB1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32572 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 500 Dilution and a 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.

All lanes:

Western blot - Anti-beta Catenin antibody [E247] - ChIP Grade (<a href='/en-us/products/primary-antibodies/beta-catenin-antibody-e247-chip-grade-ab32572'>ab32572</a>) at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

CTNNB1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human CTNNB1 (beta Catenin) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-ctnnb1-beta-catenin-knockout-hela-cell-line-ab255352'>ab255352</a>)

Predicted band size: 85 kDa

Observed band size: 86 kDa

false

ChIC/CUT&RUN sequencing - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)
  • ChIC/CUT&RUN-seq

Supplier Data

ChIC/CUT&RUN sequencing - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)

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

CUT&RUN profiling with B-Catenin antibody demonstrates robust genome-wide enrichment in cells. Heatmaps of genome-wide signal flanking annotated transcription start sites (TSSs, +/- 2 kbp) display CUT&RUN data generated using the CUTANA™ CUT&RUN Kit (EpiCypher 14-1048) with B-Catenin antibody (Abcam ab32572, 0.5 µg). 500,000 HeLa cells were used per reaction. IgG antibody was included as a negative control to assess non-specific background. Libraries were prepared using the CUTANA™ CUT&RUN Library Prep Kit (EpiCypher 14-1001). Sequencing was performed with paired-end 50 bp reads, and data were processed on CUTANA™ Cloud (cloud.epicypher.com) by alignment to the hg38 genome. Heatmaps were generated using ChAsE (Younesy et al., Bioinformatics 2016; PMID 27378294). Row-linked data are ranked by intensity relative to B-Catenin, with red indicating high localized enrichment and blue denoting background.

ChIC/CUT&RUN sequencing - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)
  • ChIC/CUT&RUN-seq

Supplier Data

ChIC/CUT&RUN sequencing - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)

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

CUT&RUN profiling with B-Catenin antibody reveals the expected genomic enrichment pattern in cells. Representative genome browser tracks show CUT&RUN data generated using the CUTANA™ CUT&RUN Kit (EpiCypher 14-1048) with B-Catenin antibody (Abcam ab32572, 0.5 µg). 500,000 HeLa cells were used per reaction. IgG, H3K4me3, and H3K27me3 antibodies were included as controls to assess non-specific background, active promoters, and repressed chromatin, respectively. Libraries were prepared using the CUTANA™ CUT&RUN Library Prep Kit (EpiCypher 14-1001). Sequencing was performed with paired-end 50 bp reads, and data were processed on CUTANA™ Cloud (cloud.epicypher.com) by alignment to the hg38 genome. Images were generated using Integrative Genomics Viewer (IGV, Broad Institute).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)
  • IHC-P

AbReview36286****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)

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).

This image is courtesy of an anonymous Abreview.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)
  • IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - BSA and Azide free (AB196204)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32572). Tissue Microarrays stained for Anti-beta Catenin antibody [E247] - ChIP Grade using ab32572 in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negaive (cross mark) staining per sample type tested. The section was incubated with ab32572 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

E247

Isotype

IgG

Carrier free

Yes

Reacts with

Mouse, Rat, Human

Applications

ChIC/CUT&RUN-seq, IHC-P, ICC/IF, mIHC, IP, WB, ChIP

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Specificity

This antibody is not suitable for ICC testing in mouse and rat species.

Our inhouse testing indicated that this antibody does not work in Raw264.7 cell line in western blot. We have an alternative antibody ab68183 detecting weak band in lower expressor Raw264.7.

Reactivity data

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Product details

What is this antibody validated in?
Anti-beta Catenin antibody [E247] - BSA and Azide free (ab196204) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Immunoprecipitation (IP), Immunohistochemistry (IHC-P), Immunocytochemistry/immunofluorescence (ICC/IF), ChIP in Human, Mouse, Rat samples.

What is the molecular weight of beta Catenin?
Anti-beta Catenin [E247] - BSA and Azide free (ab196204) specifically detects a band for beta Catenin (UniProt: P35222) at a molecular weight of 85kDa.

Trusted by the scientific community
Anti-beta Catenin [E247] - BSA and Azide free (ab196204) was first used in a scientific publication in 2016 and has been cited over 30 times in peer-reviewed journals.

Specificity confirmed
The specificity of Anti-beta Catenin antibody [E247] - BSA and Azide free (ab196204) has been confirmed by Western blot testing in CTNNB1 Knockout HeLa cell line, ab255352.

Other related products
We have a range of other formats of antibody clone [E247] also available for your convenience: ab32572, Alexa Fluor® 488 - ab194118, Alexa Fluor® 647 - ab194119, HRP - ab194120, Carrier free - ab196204, Alexa Fluor® 594 - ab201771, Alexa Fluor® 555 - ab202496

Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

Properties and storage information

Form
Liquid
Purification technique
Affinity purification Protein A
Storage buffer
pH: 7.2 - 7.4 Constituents: PBS
Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Beta Catenin also known by names such as CTNNB1 or beta-chip is an important protein involved in cell signaling and adhesion. This protein has a molecular weight of around 88 kDa. Beta Catenin is expressed in many cell types and tissues indicating its widespread role in various biological processes. It functions mechanically by mediating the linkage between cadherins and the actin cytoskeleton facilitating cell-cell adhesion. Beta Catenin is also a central part of transcription regulation processes in the nucleus.
Biological function summary

This protein plays roles in both cell adhesion and the regulation of gene expression. Beta Catenin is a critical component of the Wnt signaling pathway where it can form complexes with other proteins to influence gene transcription. In the absence of Wnt signaling beta Catenin levels are low due to its degradation. However when the pathway is active it accumulates in the cytoplasm and eventually translocates to the nucleus where it interacts with TCF/LEF transcription factors to regulate the expression of target genes.

Pathways

Beta Catenin plays a central role in the Wnt signaling pathway and influences cell fate decisions and cellular proliferation. It acts in concert with proteins such as Dishevelled (DVL) and Axin to coordinate these important biological processes. In the absence of Wnt signaling proteins such as APC and GSK-3Β are responsible for beta Catenin degradation keeping its cellular levels in check. Beta Catenin's interaction with transcription factors in the nucleus makes it pivotal in the regulation of cell and tissue homeostasis.

Beta Catenin has associations with colorectal cancer and hepatocellular carcinoma. Its dysregulation can lead to unchecked cell proliferation and tumorigenesis. Often mutations in the beta Catenin gene (CTNNB1) or components of the Wnt pathway like APC are implicated in the development of these cancers. Its interplay with E-cadherin is important for maintaining tissue architecture and disruptions can lead to invasive cancer phenotypes. Understanding beta Catenin's role provides insights into therapeutic strategies for these cancers.

Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:

Target data

Key downstream component of the canonical Wnt signaling pathway (PubMed : 17524503, PubMed : 18077326, PubMed : 18086858, PubMed : 18957423, PubMed : 21262353, PubMed : 22155184, PubMed : 22647378, PubMed : 22699938). In the absence of Wnt, forms a complex with AXIN1, AXIN2, APC, CSNK1A1 and GSK3B that promotes phosphorylation on N-terminal Ser and Thr residues and ubiquitination of CTNNB1 via BTRC and its subsequent degradation by the proteasome (PubMed : 17524503, PubMed : 18077326, PubMed : 18086858, PubMed : 18957423, PubMed : 21262353, PubMed : 22155184, PubMed : 22647378, PubMed : 22699938). In the presence of Wnt ligand, CTNNB1 is not ubiquitinated and accumulates in the nucleus, where it acts as a coactivator for transcription factors of the TCF/LEF family, leading to activate Wnt responsive genes (PubMed : 17524503, PubMed : 18077326, PubMed : 18086858, PubMed : 18957423, PubMed : 21262353, PubMed : 22155184, PubMed : 22647378, PubMed : 22699938). Also acts as a coactivator for other transcription factors, such as NR5A2 (PubMed : 22187462). Promotes epithelial to mesenchymal transition/mesenchymal to epithelial transition (EMT/MET) via driving transcription of CTNNB1/TCF-target genes (PubMed : 29910125). Involved in the regulation of cell adhesion, as component of an E-cadherin : catenin adhesion complex (By similarity). Acts as a negative regulator of centrosome cohesion (PubMed : 18086858). Involved in the CDK2/PTPN6/CTNNB1/CEACAM1 pathway of insulin internalization (PubMed : 21262353). Blocks anoikis of malignant kidney and intestinal epithelial cells and promotes their anchorage-independent growth by down-regulating DAPK2 (PubMed : 18957423). Disrupts PML function and PML-NB formation by inhibiting RANBP2-mediated sumoylation of PML (PubMed : 22155184). Promotes neurogenesis by maintaining sympathetic neuroblasts within the cell cycle (By similarity). Involved in chondrocyte differentiation via interaction with SOX9 : SOX9-binding competes with the binding sites of TCF/LEF within CTNNB1, thereby inhibiting the Wnt signaling (By similarity). Acts as a positive regulator of odontoblast differentiation during mesenchymal tooth germ formation, via promoting the transcription of differentiation factors such as LEF1, BMP2 and BMP4 (By similarity). Activity is repressed in a MSX1-mediated manner at the bell stage of mesenchymal tooth germ formation which prevents premature differentiation of odontoblasts (By similarity).
See full target information CTNNB1

Publications (34)

Recent publications for all applications. Explore the full list and refine your search

Journal of cellular and molecular medicine 28:e18546 PubMed39046458

2024

The role of miR-128 and MDFI in cardiac hypertrophy and heart failure: Mechanistic.

Applications

Unspecified application

Species

Unspecified reactive species

Sun Yanjun,Gu Yunfen,Yao Haoyi,Wang Zhe,Qiu Jiapei

Frontiers in oncology 12:794101 PubMed35251971

2022

Establishment of a Novel Mouse Hepatocellular Carcinoma Model for Dynamic Monitoring of Tumor Development by Bioluminescence Imaging.

Applications

Unspecified application

Species

Unspecified reactive species

Xiangyi Cao,Yulong Zhang,Qianqian Zhou,Sujing Sun,Minwei He,Xiaohui Wang,Ping Ma,Xiaoang Yang,Liping Lv,Linsheng Zhan

Organogenesis 17:27-39 PubMed34965835

2021

Upregulation of miR-128 Mediates Heart Injury by Activating Wnt/β-catenin Signaling Pathway in Heart Failure Mice.

Applications

Unspecified application

Species

Unspecified reactive species

Jing-Yao Li,Xin-Chang Li,Yu-Long Tang

International journal of biological sciences 17:1289-1301 PubMed33867846

2021

Enforced expression of miR-92b blunts lipopolysaccharide-mediated inflammatory injury by activating the PI3K/AKT/β-catenin pathway via targeting PTEN.

Applications

Unspecified application

Species

Unspecified reactive species

Kangfeng Jiang,Jing Yang,Chunlian Song,Fengping He,Liangyu Yang,Xiaobing Li

Experimental biology and medicine (Maywood, N.J.) 240:45-57 PubMed25053781

2014

Featured article: induction of heme oxygenase with hemin improves pericardial adipocyte morphology and function in obese Zucker rats by enhancing proteins of regeneration.

Applications

Unspecified application

Species

Rat

Joseph Fomusi Ndisang,Shuchita Tiwari

International journal of medical sciences 11:971-8 PubMed25076852

2014

Wnt signaling as a possible promoting factor of cell differentiation in pleomorphic adenomas.

Applications

IHC-P

Species

Human

Yukiko Okuda,Keisuke Nakano,Koji Suzuki,Yoshihiko Sugita,Katsutoshi Kubo,Hatsuhiko Maeda,Norimasa Okafuji,Hiromasa Hasegawa,Toshiyuki Kawakami

PloS one 9:e99137 PubMed24918446

2014

Dose-dependent effect of estrogen suppresses the osteo-adipogenic transdifferentiation of osteoblasts via canonical Wnt signaling pathway.

Applications

Unspecified application

Species

Mouse

Bo Gao,Qiang Huang,Yan-Shui Lin,Bo-Yuan Wei,Yun-Shan Guo,Zhen Sun,Long Wang,Jing Fan,Hong-Yang Zhang,Yue-Hu Han,Xiao-Jie Li,Jun Shi,Jian Liu,Liu Yang,Zhuo-Jing Luo

PloS one 9:e93364 PubMed24806734

2014

Wnt/β-catenin pathway regulates cementogenic differentiation of adipose tissue-deprived stem cells in dental follicle cell-conditioned medium.

Applications

WB, ICC/IF

Species

Rat, Rat

Na Liu,Bin Gu,Ning Liu,Xin Nie,Bo Zhang,Xia Zhou,Manjing Deng

Oncology letters 8:85-90 PubMed24959224

2014

Bone marrow mesenchymal stem cell-derived Wnt5a inhibits leukemia cell progression via activation of the non-canonical Wnt signaling pathway.

Applications

WB

Species

Unspecified reactive species

Ya Li Shen,Qing Luo,Yu Xia Guo,Gai Huai Zheng,Jie Yu,You Hua Xu

PloS one 9:e91086 PubMed24755605

2014

Wnt signaling in form deprivation myopia of the mice retina.

Applications

WB

Species

Mouse

Mingming Ma,Zhengwei Zhang,Ergang Du,Wenjing Zheng,Qing Gu,Xun Xu,Bilian Ke
View all publications

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