Recombinant Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] - BSA and Azide free (ab305262)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR26155-110-1] to beta Catenin - BSA and Azide free
- Suitable for: WB, Dot blot, Flow Cyt (Intra), IHC-P, IP, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] - BSA and Azide free
See all beta Catenin primary antibodies -
Description
Rabbit monoclonal [EPR26155-110-1] to beta Catenin - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: WB, Dot blot, Flow Cyt (Intra), IHC-P, IP, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Wild-type HAP1 whole cell lysate. HeLa, HEK-293T, NIH/3T3 and PC-12 whole cell lysate. IHC-P: Human, rat and mouse colon tissue. Human ovarian cancer tissue. Wild-type HAP1 pellet. ICC/IF: Wild-type HAP1 cells. NIH/3T3 cells. Flow Cyt (Intra): Wild-type HAP1 cells. NIH/3T3 cells. IP: HeLa and NIH/3T3 whole cell lysate. Dot: beta Catenin non-phospho peptide, beta Catenin phospho (S33), (S33/S37), (S33/S37/T41), (S37), (S37/T41), (T41), (S33/T41) peptides.
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General notes
AB305262 is the carrier-free version of AB305261.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
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.
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.
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. -
Storage buffer
pH: 7.20
Constituent: 100% PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR26155-110-1 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab305262 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
Use at an assay dependent concentration.
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Dot blot |
Use at an assay dependent concentration.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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IP |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
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Notes |
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WB
Use at an assay dependent concentration. |
Dot blot
Use at an assay dependent concentration. |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
IP
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
Target
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Function
Key dowstream component of the canonical Wnt signaling pathway. 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. 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.
Involved in the regulation of cell adhesion. The majority of beta-catenin is localized to the cell membrane and is part of E-cadherin/catenin adhesion complexes which are proposed to couple cadherins to the actin cytoskeleton. -
Tissue specificity
Expressed in several hair follicle cell types: basal and peripheral matrix cells, and cells of the outer and inner root sheaths. Expressed in colon. -
Involvement in disease
Defects in CTNNB1 are associated with colorectal cancer (CRC) [MIM:114500].
Note=Activating mutations in CTNNB1 have oncogenic activity resulting in tumor development. Somatic mutations are found in various tumor types, including colon cancers, ovarian and prostate carcinomas, hepatoblastoma (HB), hepatocellular carcinoma (HCC). HBs are malignant embryonal tumors mainly affecting young children in the first three years of life.
Defects in CTNNB1 are a cause of pilomatrixoma (PTR) [MIM:132600]; a common benign skin tumor.
Defects in CTNNB1 are a cause of medulloblastoma (MDB) [MIM:155255]. MDB is a malignant, invasive embryonal tumor of the cerebellum with a preferential manifestation in children.
Defects in CTNNB1 are a cause of susceptibility to ovarian cancer (OC) [MIM:167000]. Ovarian cancer common malignancy originating from ovarian tissue. Although many histologic types of ovarian neoplasms have been described, epithelial ovarian carcinoma is the most common form. Ovarian cancers are often asymptomatic and the recognized signs and symptoms, even of late-stage disease, are vague. Consequently, most patients are diagnosed with advanced disease.
Note=A chromosomal aberration involving CTNNB1 is found in salivary gland pleiomorphic adenomas, the most common benign epithelial tumors of the salivary gland. Translocation t(3;8)(p21;q12) with PLAG1. -
Sequence similarities
Belongs to the beta-catenin family.
Contains 12 ARM repeats. -
Post-translational
modificationsPhosphorylation by GSK3B requires prior phosphorylation of Ser-45 by another kinase. Phosphorylation proceeds then from Thr-41 to Ser-37 and Ser-33.
EGF stimulates tyrosine phosphorylation. Phosphorylation on Tyr-654 decreases CDH1 binding and enhances TBP binding.
Ubiquitinated by the SCF(BTRC) E3 ligase complex when phosphorylated by GSK3B, leading to its degradation. Ubiquitinated by a E3 ubiquitin ligase complex containing UBE2D1, SIAH1, CACYBP/SIP, SKP1, APC and TBL1X, leading to its subsequent proteasomal degradation. -
Cellular localization
Cytoplasm. Nucleus. Cytoplasm > cytoskeleton. Cell junction > adherens junction. Cell junction. Cell membrane. Cytoplasmic when it is unstabilized (high level of phosphorylation) or bound to CDH1. Translocates to the nucleus when it is stabilized (low level of phosphorylation). Interaction with GLIS2 and MUC1 promotes nuclear translocation. Interaction with EMD inhibits nuclear localization. - Information by UniProt
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Database links
- Entrez Gene: 1499 Human
- Entrez Gene: 12387 Mouse
- Entrez Gene: 84353 Rat
- Omim: 116806 Human
- SwissProt: P35222 Human
- SwissProt: Q02248 Mouse
- SwissProt: Q9WU82 Rat
- Unigene: 476018 Human
see all -
Alternative names
- b-catenin antibody
- Beta catenin antibody
- Beta-catenin antibody
see all
Images
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All lanes : Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] (ab305261) at 1/1000 dilution
Lane 1 : Wild-type HAP1(human chronic myelogenous leukemia near-haploid cell line) whole cell lysate
Lane 2 : CTNNB1 (beta Catenin) knockout HAP1 whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (ab216773) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (ab216776) at 1/10000 dilution
Observed band size: 95 kDa why is the actual band size different from the predicted?This data was developed using ab305261, the same antibody clone in a different buffer formulation.
Blocking / Diluting buffer and concentration:
Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS
The samples were run on a Bis-Tris gel.
Performed under reducing conditions.
False colour image of Western blot: Anti-beta Catenin non-phospho (active) S45 antibody (ab305261) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody 6C5 (ab8245) loading control staining at 1/20000 dilution, shown in red.
In Western blot, ab305261 was shown to bind specifically to beta Catenin non-phospho (active) S45. A band was observed at 95 kDa in wild-type HAP1 cell lysates with no signal observed at this size in CTNNB1 (beta Catenin) knockout cell line. To generate this image, wild-type and CTNNB1 (beta Catenin) knockout HAP1 cell lysates were analyzed. First, samples were run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in in Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS 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. -
Lane 1 : Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] - BSA and Azide free (ab305262) at 1/1000 dilution
Lanes 2-4 : Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] (ab305261) at 1/1000 dilution
Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : 293T (human embryonic kidney epithelial cell) whole cell lysate
Lane 3 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lane 4 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Observed band size: 95 kDa why is the actual band size different from the predicted?This data was developed using ab305261, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Lysates were freshly made and used for Western Blotting immediately to minimize protein degradation.
Exposure time: 180 seconds
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Lane 1: beta Catenin non-phospho peptide
Lane 2: beta Catenin phospho (S33) peptide
Lane 3: beta Catenin phospho (S33/S37) peptide
Lane 4: beta Catenin phospho (S33/S37/T41) peptide
Lane 5: beta Catenin phospho (S37) peptide
Lane 6: beta Catenin phospho (S37/T41) peptide
Lane 7: beta Catenin phospho (T41) peptide
Lane 8: beta Catenin phospho (S33/T41) peptide
Lane 9: beta Catenin phospho (S33/S37/T41/S45) peptide
Lane 10: beta Catenin phospho (S37/T41/S45) peptide
Lane 11: beta Catenin phospho (T41/S45) peptide
Lane 12: beta Catenin phospho (S45) peptide
Lane 13: beta Catenin phospho (S33/T41/S45) peptide
Lane 14: beta Catenin phospho (S33/S45) peptide
Lane 15: beta Catenin phospho (S37/S45) peptide
This data was developed using ab305261, the same antibody clone in a different buffer formulation.
Dot blot analysis of beta Catenin non-phospho (active) S45 using ab305261 at 1/1000 dilution (0.454 µg/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100,000 dilution.
Exposure time: 180 seconds
Blocking and diluting buffer and concentration: 5% NFDM/TBST
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This data was developed using ab305261, the same antibody clone in a different buffer formulation.
beta Catenin non-phospho (active) S45 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 µg with ab305261 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab305261 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 µg
Lane 2: ab305261 IP in NIH/3T3 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab305261 in NIH/3T3 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 50 seconds
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This data was developed using ab305261, the same antibody clone in a different buffer formulation.
beta Catenin non-phospho (active) S45 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg with ab305261 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab305261 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg
Lane 2: ab305261 IP in HeLa whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab305261 in HeLa whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 50 seconds
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This data was developed using ab305261, the same antibody clone in a different buffer formulation.
Flow cytometric (Intracellular) analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell, Right) / CTNNB1 (beta Catenin) knockout HAP1 (Left) cells labelling beta Catenin non-phospho (active) S45 with ab305261 at 1/500 dilution (0.1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.
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This data was developed using ab305261, the same antibody clone in a different buffer formulation.
Flow cytometric (Intracellular) analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling beta Catenin non-phospho (active) S45 with ab305261 at 1/50 dilution (1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.
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This data was developed using ab305261, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling beta Catenin non-phospho (active) S45 with ab305261 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing membranous and weak cytoplasmic staining in NIH/3T3 cell line. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.
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This data was developed using ab305261, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized CTNNB1 (beta Catenin) knockout HAP1 cells labelling beta Catenin non-phospho (active) S45 with ab305261 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in parental HAP1 cell line, and no staining in CTNNB1 (beta Catenin) knockout HAP1 cell line. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.
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This data was developed using ab305261, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Wild-type HAP1 (Panel A) positive staining labeling beta Catenin non-phospho (active) S45 with ab305261 at 1/2000 (0.227 µg/ml) followed by a ready to use Leica DS9800 (Bond™ Polymer Refine Detection) kit. No staining on CTNNB1 (beta Catenin) knockout HAP1 (Panel B). The section was incubated with ab305261 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Leica DS9800 (Bond™ Polymer Refine Detection) kit.
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins
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This data was developed using ab305261, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling beta Catenin non-phospho (active) S45 with ab305261 at 1/2000 (0.227 µg/ml) followed by a ready to use Leica S9800 (Bond™ Polymer Refine Detection) kit. Positive staining on rat colon. The section was incubated with ab305261 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Leica DS9800 (Bond™ Polymer Refine Detection) kit.
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins
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This data was developed using ab305261, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling beta Catenin non-phospho (active) S45 with ab305261 at 1/2000 (0.227 µg/ml) followed by a ready to use Leica DS9800 (Bond™ Polymer Refine Detection) kit. Positive staining on mouse colon. The section was incubated with ab305261 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Leica DS9800 (Bond™ Polymer Refine Detection) was used.
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins
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This data was developed using ab305261, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human ovarian cancer tissue labeling beta Catenin non-phospho (active) S45 with ab305261 at 1/500 (0.908 µg/ml) followed by a ready to use Leica DS9800 (Bond™ Polymer Refine Detection) kit. Positive staining on human ovarian cancer. The section was incubated with ab305261 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Leica DS9800 (Bond™ Polymer Refine Detection) kit.
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins
-
This data was developed using ab305261, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human colon tissue labeling beta Catenin non-phospho (active) S45 with ab305261 at 1/500 (0.908 µg/ml) followed by a ready to use Leica DS9800 (Bond™ Polymer Refine Detection) kit. Positive staining on human colon. The section was incubated with ab305261 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Leica DS9800 (Bond™ Polymer Refine Detection) kit.
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab305262 has not yet been referenced specifically in any publications.