WB
Western blot analysis of Beta Catenin on different lysates with Rabbit anti-Beta Catenin antibody at 1/1,000 dilution. Lane 1: NIH/3T3 cell lysate (10 µg/Lane), Lane 2: Rat brain tissue lysate (20 µg/Lane), 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 at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:100,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-Beta Catenin antibody at 1/1,000 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 at 1/1,000 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.ICC/IF
Immunocytochemistry analysis of C6 cells labeling Beta Catenin with Rabbit anti-Beta Catenin antibody 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 at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.IF-P
Immunofluorescence analysis of paraffin-embedded mouse colon tissue labeling Beta Catenin with Rabbit anti-Beta Catenin antibody 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 (green) at 1/100 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).IF-F
Immunofluorescence analysis of frozen mouse colon tissue with Rabbit anti-Beta Catenin antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. 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 (green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).FC
Flow cytometric analysis of A431 cells labeling Beta Catenin. Cells were fixed and permeabilized. Then stained with the primary antibody ( 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
Beta Catenin was immunoprecipitated from 0.2 mg rat brain tissue lysate with Rabbit anti-Beta Catenin antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Rabbit anti-Beta Catenin antibody at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: Rat brain tissue lysate (input), Lane 2: Rabbit anti-Beta Catenin antibody IP in rat brain tissue lysate, Lane 3: Rabbit IgG instead of Rabbit anti-Beta Catenin antibody in rat brain tissue lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 2 seconds.| Product Name | Beta Catenin Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | β-Catenin is a key downstream effector in the Wnt signaling pathway. It is implicated in two major biological processes in vertebrates: early embryonic development and tumorigenesis. Phosphorylation at Ser-552 by AMPK promotes stabilizion of the protein, enhancing TCF/LEF-mediated transcription. Phosphorylation on Tyr-654 decreases CDH1 binding and enhances TBP binding. GSK-3β destabilizes β-catenin by phosphorylating it at Ser33, Ser37, and Thr41. Mutations at these sites result in the stabilization of β-catenin protein levels and have been found in many tumor cell lines. |
| Immunogen | Synthetic peptide within human Beta-Catenin aa 30-70. |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Recombinant rabbit |
| Tested Applications | FCICC/IFIF-FIF-PIHCIPWB |
| WB:1:1,000-1:2000 IHC:1:200-1:1000 ICC/IF:1:100 IF-P:1:100 IF-F:1:200 FC:1:1000 IP:1-2μg/sample |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Alternative Names | Beta catenin antibody
Beta-catenin antibody
Cadherin associated protein antibody
Catenin (cadherin associated protein) beta 1 88kDa antibody Catenin beta 1 antibody Catenin beta-1 antibody CATNB antibody CHBCAT antibody CTNB1_HUMAN antibody CTNNB antibody CTNNB1 antibody DKFZp686D02253 antibody FLJ25606 antibody FLJ37923 antibody OTTHUMP00000162082 antibody OTTHUMP00000165222 antibody OTTHUMP00000165223 antibody OTTHUMP00000209288 antibody OTTHUMP00000209289 antibody |
|---|---|
| Molecular Weight(MW) | 92 kDa |
| Cellular Localization | Cytoplasm, Nucleus, Cell membrane, Cell junction. |
WB
Western blot analysis of Beta Catenin on different lysates with Rabbit anti-Beta Catenin antibody at 1/1,000 dilution. Lane 1: NIH/3T3 cell lysate (10 µg/Lane), Lane 2: Rat brain tissue lysate (20 µg/Lane), 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 at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1:100,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-Beta Catenin antibody at 1/1,000 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 at 1/1,000 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.
ICC/IF
Immunocytochemistry analysis of C6 cells labeling Beta Catenin with Rabbit anti-Beta Catenin antibody 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 at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
IF-P
Immunofluorescence analysis of paraffin-embedded mouse colon tissue labeling Beta Catenin with Rabbit anti-Beta Catenin antibody 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 (green) at 1/100 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
IF-F
Immunofluorescence analysis of frozen mouse colon tissue with Rabbit anti-Beta Catenin antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. 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 (green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
FC
Flow cytometric analysis of A431 cells labeling Beta Catenin. Cells were fixed and permeabilized. Then stained with the primary antibody ( 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
Beta Catenin was immunoprecipitated from 0.2 mg rat brain tissue lysate with Rabbit anti-Beta Catenin antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Rabbit anti-Beta Catenin antibody at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: Rat brain tissue lysate (input), Lane 2: Rabbit anti-Beta Catenin antibody IP in rat brain tissue lysate, Lane 3: Rabbit IgG instead of Rabbit anti-Beta Catenin antibody in rat brain tissue lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 2 seconds.| Application Notes | WB:1:1,000-1:2000 IHC:1:200-1:1000 ICC/IF:1:100 IF-P:1:100 IF-F:1:200 FC:1:1000 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 1%BSA, 40%Glycerol. Preservative: 0.05% Sodium Azide. |
