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beta Catenin antibody (Catenin, beta) (N-Term) Primary Antibody

CATNB Reactivity: Human, Mouse, Rat ChIP, CUT&RUN, ICC, IF, IHC (p), IP, WB Host: Rabbit Polyclonal unconjugated
Pubmed (6) Independent Validation (1)
Catalog No. ABIN2855042
$509.08
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100 μL
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  • Target
    beta Catenin (CATNB)
    Binding Specificity
    • 46
    • 31
    • 28
    • 27
    • 22
    • 22
    • 18
    • 17
    • 16
    • 16
    • 16
    • 15
    • 10
    • 10
    • 7
    • 7
    • 3
    • 3
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    N-Term
    Reactivity
    • 376
    • 266
    • 254
    • 40
    • 26
    • 24
    • 23
    • 23
    • 6
    • 6
    • 5
    • 4
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Human, Mouse, Rat
    Host
    • 293
    • 84
    • 4
    • 2
    • 1
    Rabbit
    Clonality
    • 286
    • 98
    Polyclonal
    Conjugate
    • 140
    • 24
    • 16
    • 15
    • 15
    • 15
    • 15
    • 15
    • 11
    • 10
    • 9
    • 9
    • 9
    • 9
    • 9
    • 8
    • 4
    • 4
    • 4
    • 4
    • 4
    • 4
    • 4
    • 4
    • 4
    • 4
    • 4
    • 4
    • 4
    • 1
    • 1
    • 1
    This beta Catenin antibody is un-conjugated
    Application
    • 212
    • 194
    • 119
    • 119
    • 95
    • 93
    • 82
    • 51
    • 33
    • 32
    • 26
    • 16
    • 13
    • 1
    • 1
    Chromatin Immunoprecipitation (ChIP), Cleavage Under Targets and Release Using Nuclease (CUT&RUN), Immunocytochemistry (ICC), Immunofluorescence (IF), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunoprecipitation (IP), Western Blotting (WB)
    Cross-Reactivity
    Rhesus Monkey
    Cross-Reactivity (Details)
    Rhesus Monkey (100 %)
    Characteristics
    Rabbit Polyclonal antibody to beta Catenin (catenin (cadherin-associated protein), beta 1, 88 kDa)
    beta Catenin antibody [N1N2-2], N-term
    Purification
    Purified by antigen-affinity chromatography.
    Immunogen
    Recombinant protein encompassing a sequence within the N-terminus region of human beta Catenin. The exact sequence is proprietary.
    Isotype
    IgG
  • Application Notes
    The rabbit anti-beta Catenin antibody ABIN2855042 is suitable for use in ChIP, CUT&RUN, immunocytochemistry, immunofluorescence, immunohistochemistry, immunoprecipitation, and Western Blot. Specific conditions for each assay should be optimized by the end user. General ABIN2855042 dilution recommendations for different applications are as follows:
    • ICC/IF: 1:100-1:1,000
    • IHC (FFPE): 1:100-1:1,000
    • IP: 1:50-1:100
    • WB: 1:500-1:3,000
    • CUT&RUN: 1:100
    Comment

    Positive Control: A549 , HeLa , H1299 , HCT-116 , PC-12 , Mouse brain , *human fibroblast , *LX2

    Restrictions
    For Research Use only
  • Validation #104235 (Cleavage Under Targets and Release Using Nuclease)
    'Independent Validation' Badge
    by
    Cantù Lab, Gene Regulation during Development and Disease, Linköping University
    No.
    #104235
    Date
    06/28/2021
    Antigen
    beta Catenin
    Lot Number
    42536
    Method validated
    Cleavage Under Targets and Release Using Nuclease
    Positive Control

    Recombinant anti-H3K27me3 CUT&RUN Positive Control antibody (antibodies-online, ABIN6923144)

    Negative Control
    Monoclonal anti-FLAG (Sigma-Aldrich, F3165)
    Notes

    Passed. ABIN2855042 allows for beta Catenin-targeted digestion using CUT&RUN on HEK-293 nuclei.

    'Independent Validation' Badge
    Validation Images
    Full Methods
    Primary Antibody
    ABIN2855042 
    Secondary Antibody
    Full Protocol
    • Cell harvest
      • Harvest 500,000 HEK293T cells per antibody to be used at RT.
      • Centrifuge cell solution 3 min at 600 x g at RT.
      • Remove the liquid carefully.
      • Gently resuspend cells in 1 mL Wash Buffer (20 mM HEPES pH 7.5, 150 mM NaCl, 0.5 mM Spermidine, Roche Complete Protease Inhibitor EDTA-free) by pipetting and transfer cell solution to a 2 mL microcentrifuge tube.
      • Centrifuge cell solution 3 min at 600 x g at RT and discard the supernatant.
      • Repeat twice for a total of three washes.
      • Resuspend cell pellet in 1 mL Wash Buffer by gently pipetting.
    • Concanavalin A beads preparation
      • Prepare one 1.5 mL microcentrifuge tube.
      • Gently resuspend the magnetic Concanavalin A Beads (antibodies-online, ABIN6923139).
      • Pipette 40 µL Con A Beads slurry for each sample into the 1.5 mL microcentrifuge tube.
      • Place the tube on a magnet stand until the fluid is clear. Remove the liquid carefully.
      • Remove the microcentrifuge tube from the magnetic stand.
      • Pipette 1 mL Binding Buffer (20 mM HEPES pH 7.5, 10 mM KCl, 1 mM CaCl2, 1 mM MnCl2) into each tube and resuspend ConA beads by gentle pipetting.
      • Spin down the liquid from the lid with a quick pulse in a table-top centrifuge.
      • Place the tubes on a magnet stand until the fluid is clear. Remove the liquid carefully.
      • Remove the microcentrifuge tube from the magnetic stand.
      • Repeat twice for a total of three washes.
      • Gently resuspend the ConA Beads in a volume of Binding Buffer corresponding to the original volume of bead slurry, i.e. 40 µL per sample.
    • Nuclei immobilization – binding to Concanavalin A beads
      • Carefully vortex the cell suspension and add 10 µL of the Con A beads in Binding Buffer to the cell suspension for each sample.
      • Close tube tightly and rotate for 10 min at RT.
    • Primary antibody binding
      • oDivide nuclei suspension into separate 2 mL microcentrifuge tubes, one for each antibody (500,000 nuclei per sample).
      • Place the microcentrifuge tubes on a magnetic stand until the fluid is clear. Remove the liquid carefully.
      • Remove the microcentrifuge tubes from the magnetic stand.
      • Place each tube at a low angle on the vortex mixer set to a low speed and add 150 µL Digitonin Wash buffer (wash buffer with 0.025% (wt/vol) Digitonin) supplemented with 2 mM EDTA.
      • Gently vortex the microcentrifuge tubes until the beads are resuspended.
      • Add 1.5 µL antibody (anti-beta Catenin antibody ABIN2855042, anti-H3K27me3 positive control antibody ABIN6923144, and anti-FLAG tag antibody negative control) to the respective tube, corresponding to a 1:100 dilution.
      • Rotate the microcentrifuge tubes ON at 4 °C.
      • Spin down the liquid and place the tubes on a magnet stand until the fluid is clear. Remove the liquid carefully.
      • Remove the microcentrifuge tubes from the magnetic stand.
      • Resuspend with 1 mL Digitonin Wash Buffer and mix by inversion. If clumping occurs, gently remove the clumps with a 1 ml pipette tip.
      • Repeat once for a total of two washes.
    • pAG-MNase Binding
      • Place the tubes on a magnet stand until the fluid is clear. Remove the liquid carefully.
      • Remove the microcentrifuge tubes from the magnetic stand.
      • o Vortex the sample at low speed and add 2.5 µL (0.05 volumes) CUTANA pAG-MNase for ChIC/CUT&RUN Assays (ABIN6950951) in 50 µL Digitonin Wash Buffer per sample, gently resuspending the beads by pipetting.
      • Rotate the microcentrifuge tubes for 1 h at 4 °C.
      • Spin down the liquid and place the tubes on a magnet stand until the fluid is clear. Remove the liquid carefully.
      • Remove the microcentrifuge tubes from the magnetic stand.
      • Resuspend with 1 mL Digitonin Wash Buffer and mix by inversion. If clumping occurs, gently remove the clumps with a 1 mL pipette tip.
      • Repeat once for a total of two washes.
    • MNase digestion and release of pAG-MNase-antibody-chromatin complexes
      • Spin down the liquid from the lid with a quick pulse in a table-top centrifuge.
      • Place the tubes on a magnet stand until the fluid is clear. Remove the liquid carefully.
      • Place each tube at a low angle on the vortex mixer set to a low speed and add 100 μL Digitonin Wash buffer per sample along the side of the tube.
      • Place tubes in a heat block, kept on ice, and allow to chill.
      • Add 2 μL 0.1 M CaCl2 to each sample.
      • Incubate tubes at 0 °C for 30 min.
      • Add 100 μL 2xSTOP buffer (340 mM NaCl, 20 mM EDTA, 4 mM EGTA, 0.05% (wt/vol) Digitonin, 100 μg/mL RNAse A, 50 μg/mL Glycogen).
      • Incubate tubes at 37 °C for 30 min.
      • Place the tubes on a magnet stand until the fluid is clear.
      • Transfer the supernatant containing the pA-MNase-bound digested chromatin fragments to fresh 1.5 mL microcentrifuge tubes.
    • DNA extraction
      • Add 2 µL 10% SDS to a final concentration of 0.1% and 2.5 µL Proteinase K (20 mg/mL) to each supernatant.
      • Gently vortex tubes at a low speed of approximately 1,100 rpm.
      • Incubate tubes at 50 °C for 1 h.
      • Add 200 µL PCI to tube.
      • Vortex tubes thoroughly at high speed until the liquid appears milky.
      • Centrifuge tubes in a tabletop centrifuge at 16,000 x g at RT for 5 min.
      • Carefully transfer to upper aqueous phase to a fresh 1.5 mL microcentrifuge tube containing 2 µL glycogen (diluted 1:10 to 2 mg/mL from the 20 mg/mL stock solution).
      • Add 20 µL 3 M NaOAc pH 5.2.
      • Add 400 µL 100% ethanol.
      • Place tubes for at -20 °C ON.
      • Centrifuge tubes in a tabletop centrifuge at 16,000 x g at 4 °C for 5min.
      • Remove the liquid carefully with a pipette.
      • Wash pellet with 1ml 70% ethanol.
      • Centrifuge tubes in a tabletop centrifuge at 16,000 x g at 4 °C for 1 min.
      • Remove the liquid carefully with a pipette.
      • Air-dry the pellet, then dissolve in 30 µL 1 mM Tris-HCl, 0.1 mM EDTA.
    • Library preparation and sequencing
      • Prepare libraries using KAPA HyperPrep Kit using KAPA Dual-Indexed adapters according to protocol.
      • Sequence libraries on an Illumina NextSeq 500 sequencer, using a NextSeq 500/550 High Output Kit v2.5 (75 Cycles), 36bp PE.
    • Peak calling
      • Map reads to the GRCm38 (mm10) mouse genome using Bowtie2 with options: --local --very-sensitive- local --no-unal --no-mixed --no-discordant.
      • Call peaks using MACS2 with options -f BAMPE --keep-dup all –nomodel.
    Experimental Notes

    Peaks generated using ABIN2855074 for CUT&RUN aligned well with beta Catenin ChIP-seq tracks(Doumpas et al., 2019).

  • Format
    Liquid
    Concentration
    Lot specific
    Buffer
    1XPBS, 1 % BSA, 20 % Glycerol ( pH 7). 0.025 % ProClin 300 was added as a preservative.
    Preservative
    ProClin
    Precaution of Use
    This product contains ProClin: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
    Storage
    -20 °C
    Storage Comment
    Keep as concentrated solution. Aliquot and store at -20°C or below. Avoid multiple freeze-thaw cycles.
  • Yin, Yao, Deng, Wang, Cai, Shen: "Identification of candidate lncRNAs and circRNAs regulating WNT3/β-catenin signaling in essential hypertension." in: Aging, Vol. 12, Issue 9, pp. 8261-8288, 2020 (PubMed).

    Fang, Lin, Liang, Liang: "A novel c-Kit/phospho-prohibitin axis enhances ovarian cancer stemness and chemoresistance via Notch3-PBX1 and β-catenin-ABCG2 signaling." in: Journal of biomedical science, Vol. 27, Issue 1, pp. 42, 2020 (PubMed).

    Lin, Lee, Wu, Wu, Chen, Huang, Chang, Cheng, Chang, Ho, Tu, Ho: "Small G protein signalling modulator 2 (SGSM2) is involved in oestrogen receptor-positive breast cancer metastasis through enhancement of migratory cell adhesion via interaction with E-cadherin." in: Cell adhesion & migration, Vol. 13, Issue 1, pp. 120-137, 2019 (PubMed).

    Fang, VanCleave, Helmuth, Torres, Rickel, Wollenzien, Sun, Zeng, Zhao, Tao: "Targeting the Wnt/β-catenin pathway in human osteosarcoma cells." in: Oncotarget, Vol. 9, Issue 95, pp. 36780-36792, 2018 (PubMed).

    Ogawa, Hirohashi, Murai, Nishidate, Okita, Wang, Ikehara, Satoyoshi, Usui, Kubo, Nakastugawa, Kanaseki, Tsukahara, Kutomi, Furuhata, Hirata, Sato, Mizuguchi, Takemasa, Torigoe: "ST6GALNAC1 plays important roles in enhancing cancer stem phenotypes of colorectal cancer via the Akt pathway." in: Oncotarget, Vol. 8, Issue 68, pp. 112550-112564, 2017 (PubMed).

    Kung, Hung, Chien, Shih: "Control of the negative IRES trans-acting factor KHSRP by ubiquitination." in: Nucleic acids research, Vol. 45, Issue 1, pp. 271-287, 2017 (PubMed).

  • Target
    beta Catenin (CATNB)
    Alternative Name
    beta Catenin (CATNB Antibody Abstract)
    Synonyms
    Bfc, Catnb, Mesc, CTNNB, MRD19, armadillo, ctnnb, id:ibd2058, wu:fb73e10, wu:fi81c06, wu:fk25h01, beta-catenin, ctnnb1, CHBCAT, catenin (cadherin associated protein), beta 1, catenin beta 1, catenin (cadherin-associated protein), beta 1, catenin beta 1 L homeolog, Ctnnb1, CTNNB1, ctnnb1, ctnnb1.L
    Background
    Beta-catenin is an adherens junction protein. Adherens junctions (AJs, also called the zonula adherens) are critical for the establishment and maintenance of epithelial layers, such as those lining organ surfaces. AJs mediate adhesion between cells, communicate a signal that neighboring cells are present, and anchor the actin cytoskeleton. In serving these roles, AJs regulate normal cell growth and behavior. At several stages of embryogenesis, wound healing, and tumor cell metastasis, cells form and leave epithelia. This process, which involves the disruption and reestablishment of epithelial cell-cell contacts, may be regulated by the disassembly and assembly of AJs. AJs may also function in the transmission of the 'contact inhibition' signal, which instructs cells to stop dividing once an epithelial sheet is complete.[supplied by OMIM]

    Cellular Localization: Cytoplasm , Nucleus , cytoskeleton
    Molecular Weight
    85 kDa
    Gene ID
    1499
    Pathways
    Peptide Hormone Metabolism
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