HA750023

Beta Catenin Recombinant Rabbit Monoclonal Antibody [SA30-04]

cat.: HA750023

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, IF-Tissue, IP, IF-Cell, IHC-Fr, FC
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*PBS (pH7.4).
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 cancer tissue, mouse colon tissue, A431, C6.
Subcellular location:	Cytoplasm, Nucleus, Cell membrane, Cell junction
Recommended Dilutions: WB IHC-P IF-Tissue IP IF-Cell IHC-Fr FC	1:1,000-1:2,000 1:200-1:1,000 1:100 1-2μg/sample 1:100 1:200 1:1,000
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

Fig1: Western blot analysis of Beta Catenin on different lysates with Rabbit anti-Beta Catenin antibody (HA750023) 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 (HA750023) 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.

Fig2: Western blot analysis of Beta Catenin on different lysates with Rabbit anti-Beta Catenin antibody (HA750023) at 1/2,000 dilution.

Lane 1: THP-1-si NT cell lysate
Lane 2: THP-1-si Beta Catenin cell lysate

Lysates/proteins at 10 µg/Lane.

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

Exposure time: 3 minutes; ECL: K1801;

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 (HA750023) at 1/2,000 dilution was used in primary antibody dilution at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig3: Western blot analysis of Beta Catenin on different lysates with Rabbit anti-Beta Catenin antibody (HA750023) at 1/2,000 dilution.

Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-Beta Catenin KD cell lysate

Lysates/proteins at 10 µg/Lane.

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

Exposure time: 40 seconds; ECL: K1801;

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 (HA750023) at 1/2,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig4: Western blot analysis of Beta Catenin on different lysates with Rabbit anti-Beta Catenin antibody (HA750023) 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 (HA750023) 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.

	Fig5: Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-Beta Catenin antibody (HA750023) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 (HA750023) 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.
	Fig6: Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-Beta Catenin antibody (HA750023) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 (HA750023) 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.
	Fig7: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-Beta Catenin antibody (HA750023) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) 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 (HA750023) 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.

	Fig8: Immunofluorescence analysis of paraffin-embedded human colon cancer tissue labeling Beta Catenin with Rabbit anti-Beta Catenin antibody (HA750023) 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 (HA750023, 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).
	Fig9: Immunofluorescence analysis of paraffin-embedded mouse colon tissue labeling Beta Catenin with Rabbit anti-Beta Catenin antibody (HA750023) 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 (HA750023, 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).
	Fig10: Immunocytochemistry analysis of A431 cells labeling Beta Catenin with Rabbit anti-Beta Catenin antibody (HA750023) 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 (HA750023) 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.

	Fig11: Immunocytochemistry analysis of C6 cells labeling Beta Catenin with Rabbit anti-Beta Catenin antibody (HA750023) 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 (HA750023) 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.
	Fig12: Immunofluorescence analysis of frozen mouse colon tissue with Rabbit anti-Beta Catenin antibody (HA750023) 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 (HA750023, green) at 1/200 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).
	Fig13: Beta Catenin was immunoprecipitated from 0.2 mg rat brain tissue lysate with HA750023 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA750023 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: HA750023 IP in rat brain tissue lysate Lane 3: Rabbit IgG instead of HA750023 in rat brain tissue lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 2 seconds; ECL: K1801

Fig14: Flow cytometric analysis of A431 cells labeling Beta Catenin.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA750023, 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 (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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