Beta Catenin Recombinant Rabbit Monoclonal Antibody [SA30-04]
cat.: ET1601-5
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, IF-Tissue, IP, mIHC
Clonality: Monoclonal
Clone number: SA30-04
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 85 kDa
Isotype: IgG
Immunogen: Synthetic peptide within human Beta-Catenin aa 30-70.
Positive control: SW480 cell lysate, A431 cell lysate, HT-29 cell lysates, NIH/3T3 cell lysate, rat brain tissue lysate, mouse pancreas, mouse liver, human colon carcinoma tissue, mouse colon tissue.
Subcellular location: Cytoplasm, Nucleus, Cell membrane, Cell junction
Recommended Dilutions:
  WB
  IHC-P
  IF-Tissue
  IP
  mIHC
1:1,000-1:2,000
1:20-1:200
1:100
1-2μg/sample
1:2,000
Uniprot #: SwissProt: P35222 Human | Q02248 Mouse | Q9WU82 Rat
Alternative names: β Catenin Beta catenin Beta-catenin Cadherin associated protein Catenin (cadherin associated protein), beta 1, 88kDa Catenin beta 1 Catenin beta-1 CATNB CHBCAT CTNB1_HUMAN CTNNB CTNNB1 DKFZp686D02253 FLJ25606 FLJ37923 OTTHUMP00000162082 OTTHUMP00000165222 OTTHUMP00000165223 OTTHUMP00000209288 OTTHUMP00000209289
Images
ET1601-5_1.jpg Fig1: Western blot analysis of Beta Catenin on different lysates with Rabbit anti-Beta Catenin antibody (ET1601-5) at 1/2,000 dilution.

Lane 1: SW480 cell lysate
Lane 2: A431 cell lysate
Lane 3: HT-29 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 85 kDa
Observed band size: 100 kDa

Exposure time: 3 minutes;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1601-5) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.
ET1601-5_2.jpg Fig2: Western blot analysis of Beta Catenin on different lysates with Rabbit anti-Beta Catenin antibody (ET1601-5) at 1/1,000 dilution.

Lane 1: NIH/3T3 cell lysate (10 µg/Lane)
Lane 2: Rat brain tissue lysate (20 µg/Lane)

Predicted band size: 85 kDa
Observed band size: 85 kDa

Exposure time: 30 seconds;

6% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1601-5) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.
ET1601-5_3.jpg Fig3: Fluorescence multiplex immunohistochemical analysis of mouse pancreas (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-β-catenin (ET1601-5, Red), anti-Glucagon (ET1702-20, Green), anti-Insulin (ET1601-12, White), anti-CK19 (ET1601-6, Magenta) and anti-aSMA (ET1607-53, Yellow) on mouse pancreas. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKit™MH010101, www.luminiris.cn). The section was incubated in five rounds of staining: in the order of ET1601-5 (1/2,000 dilution), ET1702-20 (1/6,000 dilution), ET1601-12 (1/8,000 dilution), ET1601-6 (1/5,000 dilution) and ET1607-53 (1/10,000 dilution) for 20 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) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.
ET1601-5_4.jpg Fig4: Fluorescence multiplex immunohistochemical analysis of mouse liver (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-β-catenin (ET1601-5, Tangerine), anti-αSMA (ET1607-53, Yellow), anti-SOX9 (ET1611-56, Green), anti-Albumin (ET1702-55, Cyan) anti-GS (EM1902-39, Magenta) and anti-CK19 (ET1601-6, Orange) on mouse liver. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKit™MH010101, www.luminiris.cn). The section was incubated in six rounds of staining: in the order of ET1601-5 (1/2,000 dilution), ET1607-53 (1/3,000 dilution), ET1611-56 (1/1,500 dilution), ET1702-55 (1/3,000 dilution), EM1902-39 (1/2,000 dilution) and ET1601-6 (1/3,000 dilution) for 20 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) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.
ET1601-5_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-Beta Catenin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1601-5, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1601-5_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse colon tissue using anti-Beta Catenin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1601-5, 1/20) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1601-5_7.jpg Fig7: Immunofluorescence analysis of paraffin-embedded human colon carcinoma tissue labeling Beta Catenin with Rabbit anti-Beta Catenin antibody (ET1601-5) at 1/100 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1601-5, green) at 1/100 dilution overnight at 4 ℃, washed with PBS.

Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
ET1601-5_8.jpg Fig8: Immunofluorescence analysis of paraffin-embedded mouse colon tissue labeling Beta Catenin with Rabbit anti-Beta Catenin antibody (ET1601-5) at 1/100 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1601-5, green) at 1/100 dilution overnight at 4 ℃, washed with PBS.

Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
ET1601-5_9.jpg Fig9: Immunocytochemistry analysis of A431 cells labeling Beta Catenin with Rabbit anti-Beta Catenin antibody (ET1601-5) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Beta Catenin antibody (ET1601-5) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET1601-5_10.jpg Fig10: Immunocytochemistry analysis of C6 cells labeling Beta Catenin with Rabbit anti-Beta Catenin antibody (ET1601-5) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Beta Catenin antibody (ET1601-5) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET1601-5_11.jpg Fig11: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-Beta Catenin antibody (ET1601-5) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1601-5) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1601-5_12.jpg Fig12: Beta Catenin was immunoprecipitated from 0.2 mg rat brain tissue lysate with ET1601-5 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using ET1601-5 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: Rat brain tissue lysate (input)
Lane 2: ET1601-5 IP in rat brain tissue lysate
Lane 3: Rabbit IgG instead of ET1601-5 in rat brain tissue lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 2 seconds; ECL: K1801
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.