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

24 product images

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  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)

  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. Anti-beta Catenin antibody [E247] - ChIP Grade 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 (Anti-beta Catenin antibody [E247] - ChIP Grade ab32572).

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

  This data was developed using the same antibody in a different buffer formulation (Anti-beta Catenin antibody [E247] - ChIP Grade 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 Anti-beta Catenin antibody [E247] - ChIP Grade ab32572 (red), or 5 μg of rabbit normal IgG Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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

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  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] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-beta Catenin antibody [E247] - ChIP Grade 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 (Human CTNNB1 (beta Catenin) knockout Hep G2 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed 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

  Performed under reducing conditions.

  Predicted band size: 85 kDa

  Observed band size: 85 kDa

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  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 Alexa Fluor® 647 Anti-beta Catenin antibody [E247] ab194119 for protocol details.

  Alexa Fluor® 647 Anti-beta Catenin antibody [E247] 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 Alexa Fluor® 647 Anti-beta Catenin antibody [E247] ab194119 at 1/500 dilution (shown in red) and Alexa Fluor® 488 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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).

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  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 Alexa Fluor® 488 Anti-beta Catenin antibody [E247] ab194118 for protocol details.

  Alexa Fluor® 488 Anti-beta Catenin antibody [E247] 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 Alexa Fluor® 488 Anti-beta Catenin antibody [E247] ab194118 at 1/500 dilution (shown in green) and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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).

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  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 (Anti-beta Catenin antibody [E247] - ChIP Grade ab32572).
  

  Lanes 1- 2: Merged signal (red and green). Green - Anti-beta Catenin antibody [E247] - ChIP Grade ab32572 observed at 86 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.

  Anti-beta Catenin antibody [E247] - ChIP Grade 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 Human CTNNB1 (beta Catenin) knockout HeLa cell line ab255352 (knockout cell lysate Human CTNNB1 (beta Catenin) knockout HeLa 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. Anti-beta Catenin antibody [E247] - ChIP Grade ab32572 and Anti-GAPDH antibody [6C5] - Loading Control (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (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 (Anti-beta Catenin antibody [E247] - ChIP Grade ab32572) at 1/500 dilution

  Lane 1: Wild-type HeLa cell lysate at 20 µg

  Lane 2: CTNNB1 knockout HeLa cell lysate at 20 µg

  Performed under reducing conditions.

  Predicted band size: 85 kDa

  Observed band size: 86 kDa

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  Immunocytochemistry/ Immunofluorescence - Anti-beta Catenin antibody [E247] - BSA and Azide free (ab196204)

  Anti-beta Catenin antibody [E247] - ChIP Grade 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 Anti-beta Catenin antibody [E247] - ChIP Grade ab32572 at 1/250 dilution and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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 (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 (Anti-beta Catenin antibody [E247] - ChIP Grade ab32572).

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  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] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-beta Catenin antibody [E247] - ChIP Grade 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 Human CTNNB1 (beta Catenin II) knockout HCT116 cell line ab273712 (CRISPR-Cas9 edited cell lysate Human CTNNB1 (beta Catenin) knockout HCT116 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.

  All lanes: Western blot - Anti-beta Catenin antibody [E247] - ChIP Grade (Anti-beta Catenin antibody [E247] - ChIP Grade ab32572) 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

  Performed under reducing conditions.

  Predicted band size: 85 kDa

  Observed band size: 95 kDa

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

  Lanes 1 - 4: Merged signal (red and green). Green - Anti-beta Catenin antibody [E247] - ChIP Grade ab32572 observed at 90 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
  

  Anti-beta Catenin antibody [E247] - ChIP Grade 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. Anti-beta Catenin antibody [E247] - ChIP Grade ab32572 and Anti-GAPDH antibody [6C5] - Loading Control 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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

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  This image is courtesy of an anonymous customer review.

  Immunocytochemistry/ Immunofluorescence - Anti-beta Catenin antibody [E247] - BSA and Azide free (ab196204)

  Anti-beta Catenin antibody [E247] - ChIP Grade 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 (Anti-beta Catenin antibody [E247] - ChIP Grade ab32572).

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

  This data was developed using Anti-beta Catenin antibody [E247] - ChIP Grade ab32572, the same antibody clone in a different buffer formulation. Different batches of Anti-beta Catenin antibody [E247] - ChIP Grade 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 (Anti-beta Catenin antibody [E247] - ChIP Grade ab32572)

  Predicted band size: 85 kDa

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  This image is courtesy of an anonymous customer review.

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - BSA and Azide free (ab196204)

  Anti-beta Catenin antibody [E247] - ChIP Grade 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 (Anti-beta Catenin antibody [E247] - ChIP Grade ab32572).

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  This image is courtesy of an anonymous customer review.

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - BSA and Azide free (ab196204)

  Anti-beta Catenin antibody [E247] - ChIP Grade 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 (Anti-beta Catenin antibody [E247] - ChIP Grade ab32572).

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  Immunocytochemistry/ Immunofluorescence - Anti-beta Catenin antibody [E247] - BSA and Azide free (ab196204)

  Anti-beta Catenin antibody [E247] - ChIP Grade ab32572 staining beta Catenin in SW480 (Human colorectal adenocarcinoma cell line) cells treated with BIO (BIO, GSK3 inhibitor; cell-permeable 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 BIO, GSK3 inhibitor; cell-permeable 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 Anti-beta Catenin antibody [E247] - ChIP Grade 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 (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 (Anti-beta Catenin antibody [E247] - ChIP Grade ab32572).

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  Immunocytochemistry/ Immunofluorescence - Anti-beta Catenin antibody [E247] - BSA and Azide free (ab196204)

  Anti-beta Catenin antibody [E247] - ChIP Grade ab32572 staining beta Catenin in SK-N-SH (Human neuroblastoma cell line) cells treated with olanzapine (Olanzapine, 5-HT, dopamine, histamine and muscarinic antagonist 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 Olanzapine, 5-HT, dopamine, histamine and muscarinic antagonist 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 Anti-beta Catenin antibody [E247] - ChIP Grade 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 (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 (Anti-beta Catenin antibody [E247] - ChIP Grade ab32572).

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  This image is courtesy of an anonymous customer review.

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - BSA and Azide free (ab196204)

  Anti-beta Catenin antibody [E247] - ChIP Grade 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 (Anti-beta Catenin antibody [E247] - ChIP Grade ab32572).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - BSA and Azide free (ab196204)

  Anti-beta Catenin antibody [E247] - ChIP Grade 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 (Anti-beta Catenin antibody [E247] - ChIP Grade ab32572).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - BSA and Azide free (ab196204)

  Anti-beta Catenin antibody [E247] - ChIP Grade 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 (Anti-beta Catenin antibody [E247] - ChIP Grade ab32572).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - BSA and Azide free (ab196204)

  Anti-beta Catenin antibody [E247] - ChIP Grade 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 (Anti-beta Catenin antibody [E247] - ChIP Grade ab32572).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - BSA and Azide free (ab196204)

  Anti-beta Catenin antibody [E247] - ChIP Grade 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 (Anti-beta Catenin antibody [E247] - ChIP Grade ab32572).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - BSA and Azide free (ab196204)

  Anti-beta Catenin antibody [E247] - ChIP Grade 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 (Anti-beta Catenin antibody [E247] - ChIP Grade ab32572).

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  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

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  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] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-beta Catenin antibody [E247] - ChIP Grade 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 Human CTNNB1 knockout MCF7 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 (Anti-beta Catenin antibody [E247] - ChIP Grade ab32572).

  All lanes: Western blot - Anti-beta Catenin antibody [E247] - ChIP Grade (Anti-beta Catenin antibody [E247] - ChIP Grade ab32572) 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

  Performed under reducing conditions.

  Predicted band size: 85 kDa

  Observed band size: 85/90 kDa

  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] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-beta Catenin antibody [E247] - ChIP Grade 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 Human CTNNB1 knockout MCF7 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.

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  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 (Anti-beta Catenin antibody [E247] - ChIP Grade ab32572).
  
  Tissue Microarrays stained for Anti-beta Catenin antibody [E247] - ChIP Grade using Anti-beta Catenin antibody [E247] - ChIP Grade 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 Anti-beta Catenin antibody [E247] - ChIP Grade ab32572 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (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.