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anti-Human CTNNB1 antibody for Immunoprecipitation

Recommended CTNNB1 Antibody (supplied by: Log in to see )

Antigen
Catenin (Cadherin-Associated Protein), beta 1, 88kDa (CTNNB1) Antibodies
  • Jup
  • LOC100217813
  • CTNNB1
  • CTNNB
  • Bfc
  • Catnb
  • Mesc
  • ctnnb
  • id:ibd2058
  • wu:fb73e10
  • wu:fi81c06
  • wu:fk25h01
  • ctnnb1
  • CHBCAT
  • MRD19
  • armadillo
  • beta-catenin
  • catenin (cadherin-associated protein), beta 1, 88kDa
  • catenin (cadherin associated protein), beta 1
  • catenin (cadherin-associated protein), beta 1
  • CTNNB1
  • LOC100217813
  • Ctnnb1
  • ctnnb1
  • ctnnb1-a
  • ctnnb1-b
Reactivity
Human, Mouse (Murine), Rat (Rattus)
442
106
92
29
21
13
7
6
5
4
4
2
1
1
1
Host
Rabbit
306
154
2
2
Clonality
Polyclonal
Conjugate
This CTNNB1 antibody is conjugated to Magnetic Particles
15
12
9
6
6
6
1
Application
Immunoprecipitation (IP)
392
149
117
100
82
68
66
33
13
12
8
4
3
3
2
1
1
1
Options
Supplier
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Supplier Product No.
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Available images

Catalog No. ABIN1690141
$ 398.75
Plus shipping costs $45.00
Relevance Score ABIN Application Conjugate Host Isotype Epitope Supplier Clonality References Details
1 ABIN181394 IHC (p) IP WB Rabbit C-Term Log in to see Polyclonal 1
1 ABIN126747 EIA IF IHC (p) IP WB Mouse IgG1 Exon 3 Log in to see 9G2
1 ABIN614788 FACS IF IHC (p) IP WB Mouse IgG1 Log in to see EM-22 3
1 ABIN126748 EIA IF IHC (p) IP WB Mouse IgG2b Core Log in to see 9G10
1 ABIN126750 EIA IF IHC (p) IP WB Mouse IgG1 C-Term Log in to see 7D8
1 ABIN126746 EIA IF IHC (p) IP WB Biotin Mouse IgG1 Exon 3 Log in to see 9G2
1 ABIN126745 EIA IP WB Mouse IgG2a N-Term, Exon 2 Log in to see 7D11
1 ABIN126749 EIA IP WB Mouse IgG2a C-Term Log in to see 10H8
1 ABIN1105694 FACS IF IP WB Mouse IgG1 Log in to see 9F2 3
1 ABIN457419 ICC FACS IP WB Mouse IgG1 C-Term, AA 441-448 Log in to see EM-22 3
1 ABIN181393 IHC (p) IP WB Rabbit C-Term Log in to see Polyclonal
1 ABIN187346 IF IP WB Mouse IgG1 Log in to see 9F2
1 ABIN211079 ICC IP WB Rabbit IgG AA 29-49 Log in to see Polyclonal
1 ABIN1584165 IP IHC ELISA WB Rabbit Log in to see Polyclonal 1
1 ABIN331374 IP WB Rabbit IgG C-Term Log in to see Polyclonal
1 ABIN331369 IP WB Rabbit IgG N-Term Log in to see Polyclonal
1 ABIN604623 IP WB Sheep AA 771-781 Log in to see Polyclonal
1 ABIN574434 ICC FACS IHC (p) IP WB Mouse IgG1 Log in to see EM-22
1 ABIN386498 IHC (p) IP WB Rabbit IgG C-Term Log in to see Polyclonal
1 ABIN387410 IHC (p) IP WB Rabbit IgG C-Term Log in to see Polyclonal

Top referenced anti-CTNNB1 antibody for Immunoprecipitation

Similar anti-CTNNB1 Antibodies

Application / Reactivity Human Mouse (Murine) Rat (Rattus)
Western Blotting (WB) 377 Antibodies 97 Antibodies 83 Antibodies
RNA Interference (RNAi) 2 Antibodies
Proximity Ligation Assay (PLA) 4 Antibodies
Luminex Assay (LMNX) 8 Antibodies
Immunoprecipitation (IP) 62 Antibodies 19 Antibodies 11 Antibodies
Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)) 77 Antibodies 29 Antibodies 21 Antibodies
Immunohistochemistry (IHC) 115 Antibodies 36 Antibodies 36 Antibodies
Immunohistochemistry (Frozen Sections) (IHC (fro)) 9 Antibodies 3 Antibodies 3 Antibodies
Immunohistochemistry (Formalin-fixed Paraffin-embedded Sections) (IHC (fp)) 1 Antibodies

General

Antigen Catenin (Cadherin-Associated Protein), beta 1, 88kDa (CTNNB1) Antibodies
Reactivity Human, Mouse (Murine), Rat (Rattus)
(442), (106), (92), (29), (21), (13), (7), (6), (5), (4), (4), (2), (1), (1), (1)
Host Rabbit
(306), (154), (2), (2)
Clonality
Polyclonal
Conjugate This CTNNB1 antibody is conjugated to Magnetic Particles
(15), (12), (9), (6), (6), (6), (1)
Application Immunoprecipitation (IP)
(392), (149), (117), (100), (82), (68), (66), (33), (13), (12), (8), (4), (3), (3), (2), (1), (1), (1)
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Target Details CTNNB1

Product Details anti-CTNNB1 Antibody Application Details Handling Images back to top
Antigen
Alternative Name CTNNB1 / Beta Catenin (CTNNB1 Antibody Abstract)
Pathways WNT Signaling, Intracellular Steroid Hormone Receptor Signaling Pathway, Peptide Hormone Metabolism

Application Details

Product Details anti-CTNNB1 Antibody Target Details CTNNB1 Handling Images back to top
Restrictions For Research Use only
'Independent Validation' Badge
Antigen CTNNB1
Lot Number 416
Method validated Immunoprecipitation
Positive Control Colo205 human colorectal cell line
Notes ABIN1690141 (CTNNB1 antibody conjugated to magnetic particles) was found to be suitable for IP of beta-catenin from lysates of Colo205 cells.
Primary Antibody ABIN1690141
Protocol
  • Cell lysates:
    • Prepare detergent-free lysates of Colo205 cells on ice in low detergent 'soft' elution buffer (150mM NaCl, 50mM Tris-HCl, 0.02% Tween-20, pH8.0; see Antrobus R and Borner GH, (2011) PLoS One) supplemented with Halt Protease and Phosphatase inhibitor cocktail (Thermo Fisher, 78440, lot RE234421) using a TissueRuptor (Qiagen).
    • Pellet insoluble debris by centrifugation.
    • Transfer the supernatant and quantitate the total protein concentration by CB-X Protein Assay (G Biosciences, 786-11X, lot 152710).
  • Immunoprecipitation:
    • Dilute supernatant to 1mg/ml total protein.
    • Equilibrate CTNNB1 antibody magnetic particles (antibodies-online, ABIN1690141, lot 0416) with PBS and then wash them 3x with PBS to remove residual detergent.
    • Incubate 50µg CTNNB1 antibody magnetic particles (antibodies-online, ABIN1690141, lot 0416) with 1ml of Colo205 lysate at 1mg/ml total protein ON at 4°C on an end-over-end rotator.
    • Recover the magnetic beads on a Dynabead rack (ThermoFisher). Store the removed supernatant at -80°C for the subsequent immunoblot.
    • Wash beads 5x with 1ml ice cold PBS supplemented with Halt cocktail. Store each wash fraction at -80°C for the subsequent immunoblot.
    • Incubate immunoprecipitated proteins with 50µl 'soft' reduced detergent elution buffer (0.2% (w/v) SDS, 0.1% (v/v) Tween-20, 50mM Tris-HCl pH8.0) suitable for downstream mass spectrometric analysis.
  • Immunoblot:
    • Separate proteins on a 10% SDS-PAGE gel using a Mini-PROTEAN 3 cell electrophoresis tank (Bio-Rad, 165-3301/165-3302) at 200 V for 90min at RT and Tris-Glycine-SDS running buffer.
    • Transfer proteins to PVDF immobilon-P membrane (Millipore, IPVH00010, R6kA7917F) using a Mini Trans-Blot Electrophoretic Transfer Cell (Bio-Rad, 170-3930/170-3935) at 30V, 90mA, ON at 4°C in Tris-Glycine transfer buffer.
    • Incubate with a primary mouse purified mouse-anti-beta-catenin antibody (BD Biosciences, 610154, LOT 5113978) diluted 1:1000x in TBST (TBS, 0.1% (v/v) Tween-20) with 5% non-fat milk for 2h at RT.
    • Wash membrane 3x 5min with TBST.
    • Incubate with HRP-conjugated polyclonal goat anti-mouse secondary antibody (Dako, P0447, lot 00082889) diluted 1:2000 in TBST with 5% non-fat milk for 1h at RT.
    • Wash membrane 3x 5min with TBST.
    • Visualized immunoreactive bands by autoradiography. Incubate membrane for 1min at RT with 3ml of ECL Western Blotting Substrate (Promega, W1001, lot 0000193677) and expose to Hyperfilm ECL (Amersham, product 28906837, lot 64701) for 5min in the dark.
  • Mass spectrometric analysis:
    • Extract peptides from the gel band and submit to a tryptic digest with Sequencing Grade Modified Trypsin (Promega, V5111).
    • Run tryptic fragments on a Q Exactive (QEX) Hybrid Quadrupole-Orbitrap Mass Spectrometer (ThermoFisher).
    • Identify proteins fragments by performing an MS/MS Ion search with MASCOT (Matrix Science) against the UPR HomoSapiens 20160706 database (92,578 sequences; 36,833,215 residues).
Experimental Notes
  • ABIN1690141 proved to be be susceptible to detergents as it was evidenced by the higher efficacy of the lower detergent 'soft' elution buffer compared to RIPA buffer with PBS.
  • The IP can be improved through the use of detergent-free protein lysate preparations and by washing with PBS rather than RIPA, as strong detergents can strip beta-catenin from the antibodies on the beads. The 'soft', low detergent elution buffer formulated by Antrobus and Borner is recommended where the eluted protein is to be used for mass spectrometry.
  • Assuming that the lysate for the IP came from the same source, so there is no danger of cross-contamination, the beads can be re-used approximately four times.
Validation Images
Immunoprecipitation validation image for anti-Catenin (Cadherin-Associated Protein), beta 1, 88kDa (CTNNB1) antibody (Magnetic Particles) (ABIN1690141) A. SDS-PAGE of protein fractions from immunoprecipitation of beta-catenin with ABIN16...