Description
Mouse Catenin beta-1 (Ctnnb1) ELISA Kit
The Mouse Catenin Beta-1 (CTNNB1) ELISA Kit is specifically designed for the accurate quantification of Catenin Beta-1 levels in mouse serum, plasma, and cell culture supernatants. This kit offers high sensitivity and specificity, ensuring precise and reproducible results for a variety of research applications.Catenin Beta-1 is an important protein involved in cell adhesion and signaling pathways, playing a critical role in development, cell differentiation, and disease progression.
Its dysregulation has been associated with various diseases, including cancer, developmental disorders, and inflammatory conditions, making it a valuable biomarker for studying these processes and potential therapeutic interventions.Overall, the Mouse Catenin Beta-1 (CTNNB1) ELISA Kit provides researchers with a reliable tool for investigating the role of Catenin Beta-1 in mouse models, offering valuable insights into its function and potential implications in disease mechanisms.
| Product Name: | Mouse Catenin beta-1 (Ctnnb1) ELISA Kit |
| SKU: | MOEB0740 |
| Size: | 96T |
| Target: | Mouse Catenin beta-1 (Ctnnb1) |
| Synonyms: | Beta-catenin, Catnb |
| Assay Type: | Sandwich |
| Detection Method: | ELISA |
| Reactivity: | Mouse |
| Detection Range: | 0.156-10ng/mL |
| Sensitivity: | 0.078ng/mL |
| Intra CV: | Provided with the Kit |
| Inter CV: | Provided with the Kit |
| Linearity: | Provided with the Kit |
| Recovery: | Provided with the Kit |
| Function: | Key downstream 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. Acts as a negative regulator of centrosome cohesion. Involved in the CDK2/PTPN6/CTNNB1/CEACAM1 pathway of insulin internalization. Blocks anoikis of malignant kidney and intestinal epithelial cells and promotes their anchorage-independent growth by down-regulating DAPK2. Disrupts PML function and PML-NB formation by inhibiting RANBP2-mediated sumoylation of PML (By similarity). Promotes neurogenesis by maintaining sympathetic neuroblasts within the cell cycle (PubMed:21325504). |
| Uniprot: | Q02248 |
| Sample Type: | Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids |
| Specificity: | Natural and recombinant mouse Catenin beta-1 |
| Sub Unit: | Two separate complex-associated pools are found in the cytoplasm. The majority is present as component of an E-cadherin/ catenin adhesion complex composed of at least E-cadherin/CDH1 and beta-catenin/CTNNB1, and possibly alpha-catenin/CTNNA1; the complex is located to adherens junctions. The stable association of CTNNA1 is controversial as CTNNA1 was shown not to bind to F-actin when assembled in the complex. Alternatively, the CTNNA1-containing complex may be linked to F-actin by other proteins such as LIMA1. Binds SLC9A3R1. Interacts with PTPRU (via the cytoplasmic juxtamembrane domain) and with EMD. Interacts with SESTD1 and TRPC4. Interacts with CAV1. Interacts with PTPRJ. Interacts with PKT7. Interacts with FAT1 (via the cytoplasmic domain). Interacts with CDK2, NDRG2 and NANOS1. Interacts with NEK2 and CDK5. Interacts with CARM1, CXADR, PCDH11Y and PTK6. Interacts with RAPGEF2. Interacts with SOX7; this interaction may lead to proteasomal degradation of active CTNNB1 and thus inhibition of Wnt/beta-catenin-stimulated transcription. Identified in a complex with HINT1 and MITF. Interacts with FHIT. Interacts with FERMT2. Identified in a complex with TCF4 and FERMT2. Another cytoplasmic pool is part of a large complex containing 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. Wnt-dependent activation of DVL antagonizes the action of GSK3B. When GSK3B activity is inhibited the complex dissociates, CTNNB1 is dephosphorylated and is no longer targeted for destruction. The stabilized protein translocates to the nucleus, where it binds TCF/LEF-1 family members, TBP, BCL9, BCL9L and possibly also RUVBL1 and CHD8. Interacts with TAX1BP3 (via the PDZ domain); this interaction inhibits the transcriptional activity of CTNNB1. Interacts with AJAP1, BAIAP1 and CTNNA3. Interacts with TRPV4; the TRPV4 and CTNNB1 complex can interact with CDH1. Interacts with VCL. The CTNNB1 and TCF4 complex interacts with PML. Interacts with XIRP1. Binds CTNNBIP and EP300. CTNNB1 forms a ternary complex with LEF1 and EP300 that is disrupted by CTNNBIP1 binding. Interacts directly with AXIN1; the interaction is regulated by CDK2 phosphorylation of AXIN1. Interacts with GLIS2. Interacts with SCRIB. Interacts with TNIK and TCF7L2. Interacts with SLC30A9. Interacts with RORA. May interact with P-cadherin/CDH3. Interacts with RNF220 (By similarity). Interacts with CTNND2 (By similarity). Interacts (via the C-terminal region) with CBY1. |
| Research Area: | Cancer |
| Subcellular Location: | Cytoplasm Cytoplasm Cytoskeleton Nucleus Cell junction Adherens junction Cell membrane Cytoplasm Cytoskeleton Microtubule organizing center Centrosome Cytoplasm Cytoskeleton Spindle pole 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 promotes nuclear translocation. Interaction with EMD inhibits nuclear localization. The majority of beta-catenin is localized to the cell membrane. In interphase, colocalizes with CROCC between CEP250 puncta at the proximal end of centrioles, and this localization is dependent on CROCC and CEP250. In mitosis, when NEK2 activity increases, it localizes to centrosomes at spindle poles independent of CROCC. Colocalizes with CDK5 in the cell-cell contacts and plasma membrane of undifferentiated and differentiated neuroblastoma cells. Colocalized with RAPGEF2 and TJP1 at cell-cell contacts (By similarity). |
| Storage: | Please see kit components below for exact storage details |
| Note: | For research use only |
| UniProt Protein Function: | CTNNB1: a regulator of cell adhesion and a key downstream effector in the Wnt signaling pathway. Implicated early embryonic development and tumorigenesis. Phosphorylated and destabilized by CK1 and GSK-3beta. Stabilized cytoplasmic beta-catenin is a hallmark of a variety of cancers. Stabilized beta-catenin translocates to the nucleus, where it acts as a transcriptional activator of T-cell factor (TCF)-regulated genes. Interacts with the PDZ domain of TAX1BP3, inhibiting its transcriptional activity. Two alternatively spliced human isoforms have been described. |
| UniProt Protein Details: | Protein type:Actin-binding; Cell adhesion; Transcription factor; Oncoprotein; Motility/polarity/chemotaxis; Nuclear receptor co-regulator Cellular Component: adherens junction; apical junction complex; apical part of cell; basolateral plasma membrane; beta-catenin destruction complex; catenin complex; cell cortex; cell junction; cell projection membrane; cell-cell adherens junction; centrosome; cytoplasm; cytoskeleton; cytosol; dendritic shaft; fascia adherens; flotillin complex; focal adhesion; intercellular junction; lamellipodium; lateral plasma membrane; membrane; microvillus membrane; neuron projection; nucleus; perinuclear region of cytoplasm; plasma membrane; protein complex; synapse; tight junction; transcription factor complex; Z disc Molecular Function:alpha-catenin binding; cadherin binding; chromatin binding; DNA binding; double-stranded DNA binding; enzyme binding; estrogen receptor binding; ionotropic glutamate receptor binding; kinase binding; nitric-oxide synthase binding; nuclear hormone receptor binding; protein binding; protein C-terminus binding; protein complex binding; protein kinase binding; protein phosphatase binding; signal transducer activity; SMAD binding; transcription coactivator activity; transcription factor activity; transcription factor binding Biological Process: anterior/posterior axis specification; bone resorption; cardiac muscle cell proliferation; cell adhesion; cell differentiation; cell fate determination; cell fate specification; cell maturation; cell proliferation; cell-cell adhesion; cell-matrix adhesion; cellular morphogenesis during differentiation; cellular process; central nervous system vasculogenesis; chromatin-mediated maintenance of transcription; dorsal/ventral axis specification; dorsal/ventral pattern formation; ectoderm development; embryonic axis specification; embryonic digit morphogenesis; embryonic foregut morphogenesis; embryonic forelimb morphogenesis; embryonic heart tube development; embryonic hindlimb morphogenesis; endoderm formation; endodermal cell fate commitment; fallopian tube development; forebrain development; gastrulation with mouth forming second; genitalia morphogenesis; glial cell fate determination; hair cycle process; hair follicle morphogenesis; heart development; hemopoiesis; hindbrain development; in utero embryonic development; kidney development; layer formation in the cerebral cortex; lens morphogenesis in camera-type eye; limb development; lung development; male genitalia development; midbrain development; morphogenesis of embryonic epithelium; negative regulation of cell differentiation; negative regulation of cell proliferation; negative regulation of chondrocyte differentiation; negative regulation of mitotic cell cycle, embryonic; negative regulation of oligodendrocyte differentiation; negative regulation of osteoclast differentiation; negative regulation of protein sumoylation; negative regulation of transcription from RNA polymerase II promoter; negative regulation of transcription, DNA-dependent; nervous system development; neural plate development; neuron differentiation; neuron migration; odontogenesis of dentine-containing teeth; oocyte development; organ development; osteoclast differentiation; pancreas development; patterning of blood vessels; positive regulation of apoptosis; positive regulation of cell proliferation; positive regulation of endothelial cell differentiation; positive regulation of epithelial cell differentiation; positive regulation of fibroblast growth factor receptor signaling pathway; positive regulation of I-kappaB kinase/NF-kappaB cascade; positive regulation of MAPKKK cascade; positive regulation of mesenchymal cell proliferation; positive regulation of neuroblast proliferation; positive regulation of neuron apoptosis; positive regulation of osteoblast differentiation; positive regulation of skeletal muscle development; positive regulation of telomerase activity; positive regulation of telomere maintenance via telomerase; positive regulation of transcription factor activity; positive regulation of transcription from RNA polymerase II promoter; positive regulation of transcription, DNA-dependent; protein heterooligomerization; proximal/distal pattern formation; regulation of apoptosis; regulation of cell differentiation; regulation of cell proliferation; regulation of centriole-centriole cohesion; regulation of epithelial cell differentiation; regulation of gene expression; regulation of histone H3-K4 methylation; regulation of myelination; regulation of osteoblast differentiation; regulation of osteoclast differentiation; regulation of smooth muscle cell proliferation; regulation of T cell proliferation; regulation of transcription from RNA polymerase II promoter; regulation of transcription, DNA-dependent; response to cytokine stimulus; response to estradiol stimulus; response to estrogen stimulus; Schwann cell proliferation; skeletal development; skin development; smooth muscle cell differentiation; stem cell maintenance; synapse organization and biogenesis; synaptic transmission; synaptic vesicle transport; T cell differentiation; T cell differentiation in the thymus; thymus development; transcription, DNA-dependent; ureteric bud branching; vasculature development; vasculogenesis; Wnt receptor signaling pathway; Wnt receptor signaling pathway through beta-catenin |
| NCBI Summary: | This gene encodes not only an important cytoplasmic component of the classical cadherin adhesion complex that forms the adherens junction in epithelia and mediates cell-cell adhesion in many other tissues but also a key signaling molecule in the canonical Wnt signaling pathway that controls cell growth and differentiation during both normal development and tumorigenesis. The gene product contains a central armadillo-repeat containing domain through which it binds the cytoplasmic tail of classical cadherins; meanwhile, it also binds alpha-catenin, which further links the cadherin complex to the actin cytoskeleton either directly or indirectly. Beta-catenin is therefore necessary for the adhesive function of classical cadherins. Another key function of this protein is to mediate the canonical Wnt signaling pathway and regulate gene transcription. Without Wnt signal, cytoplasmic beta-catenin that is not associated with the cadherin complex is quickly phosphorylated at the N-terminal Ser/Thr residues by the so called degradation complex containing axin, adenomatous polyposis coli (APC), casein kinase I, and GSK3B, then ubiquitylated by beta-TrCP, and degraded by the proteasome. However, in the presence of Wnt signal, the degradation complex is disrupted and the stabilized cytoplasmic beta-catenin translocates into the nucleus, where it binds various transcription factors and, together with these factors, regulates the transcription of many downstream genes. Mutations of this gene have been linked with various types of tumors. Alternatively spliced variants have been found for this gene. [provided by RefSeq, Sep 2009] |
| UniProt Code: | Q02248 |
| NCBI GenInfo Identifier: | 399310 |
| NCBI Gene ID: | 12387 |
| NCBI Accession: | Q02248.1 |
| UniProt Secondary Accession: | Q02248,Q922W1, Q9D335, |
| UniProt Related Accession: | Q02248 |
| Molecular Weight: | 85,471 Da |
| NCBI Full Name: | Catenin beta-1 |
| NCBI Synonym Full Names: | catenin (cadherin associated protein), beta 1 |
| NCBI Official Symbol: | Ctnnb1 |
| NCBI Official Synonym Symbols: | Bfc; Mesc; Catnb |
| NCBI Protein Information: | catenin beta-1 |
| UniProt Protein Name: | Catenin beta-1 |
| UniProt Synonym Protein Names: | Beta-catenin |
| Protein Family: | Beta-catenin-like protein |
| UniProt Gene Name: | Ctnnb1 |
| UniProt Entry Name: | CTNB1_MOUSE |
| Component | Quantity (96 Assays) | Storage |
| ELISA Microplate (Dismountable) | 8×12 strips | -20°C |
| Lyophilized Standard | 2 | -20°C |
| Sample Diluent | 20ml | -20°C |
| Assay Diluent A | 10mL | -20°C |
| Assay Diluent B | 10mL | -20°C |
| Detection Reagent A | 120µL | -20°C |
| Detection Reagent B | 120µL | -20°C |
| Wash Buffer | 30mL | 4°C |
| Substrate | 10mL | 4°C |
| Stop Solution | 10mL | 4°C |
| Plate Sealer | 5 | - |
Other materials and equipment required:
- Microplate reader with 450 nm wavelength filter
- Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
- Incubator
- Deionized or distilled water
- Absorbent paper
- Buffer resevoir
*Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
Allow all reagents to reach room temperature (Please do not dissolve the reagents at 37°C directly). All the reagents should be mixed thoroughly by gently swirling before pipetting. Avoid foaming. Keep appropriate numbers of strips for 1 experiment and remove extra strips from microtiter plate. Removed strips should be resealed and stored at -20°C until the kits expiry date. Prepare all reagents, working standards and samples as directed in the previous sections. Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their experiments. We recommend running all samples in duplicate.
| Step | |
| 1. | Add Sample: Add 100µL of Standard, Blank, or Sample per well. The blank well is added with Sample diluent. Solutions are added to the bottom of micro ELISA plate well, avoid inside wall touching and foaming as possible. Mix it gently. Cover the plate with sealer we provided. Incubate for 120 minutes at 37°C. |
| 2. | Remove the liquid from each well, don't wash. Add 100µL of Detection Reagent A working solution to each well. Cover with the Plate sealer. Gently tap the plate to ensure thorough mixing. Incubate for 1 hour at 37°C. Note: if Detection Reagent A appears cloudy warm to room temperature until solution is uniform. |
| 3. | Aspirate each well and wash, repeating the process three times. Wash by filling each well with Wash Buffer (approximately 400µL) (a squirt bottle, multi-channel pipette,manifold dispenser or automated washer are needed). Complete removal of liquid at each step is essential. After the last wash, completely remove remaining Wash Buffer by aspirating or decanting. Invert the plate and pat it against thick clean absorbent paper. |
| 4. | Add 100µL of Detection Reagent B working solution to each well. Cover with the Plate sealer. Incubate for 60 minutes at 37°C. |
| 5. | Repeat the wash process for five times as conducted in step 3. |
| 6. | Add 90µL of Substrate Solution to each well. Cover with a new Plate sealer and incubate for 10-20 minutes at 37°C. Protect the plate from light. The reaction time can be shortened or extended according to the actual color change, but this should not exceed more than 30 minutes. When apparent gradient appears in standard wells, user should terminatethe reaction. |
| 7. | Add 50µL of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing. |
| 8. | Determine the optical density (OD value) of each well at once, using a micro-plate reader set to 450 nm. User should open the micro-plate reader in advance, preheat the instrument, and set the testing parameters. |
| 9. | After experiment, store all reagents according to the specified storage temperature respectively until their expiry. |
When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.
| Sample Type | Protocol |
| Serum | If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. |
| Plasma | Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit. |
| Urine & Cerebrospinal Fluid | Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid. |
| Cell culture supernatant | Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately. |
| Cell lysates | Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C. |
| Tissue homogenates | The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C. |
| Tissue lysates | Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C. |
| Breast Milk | Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles. |
