beta Catenin antibody (AA 571-781) (Alexa Fluor 488)

Catalog No. ABIN1176964

Product Details anti-beta Catenin Antibody (Alexa Fluor 488)

Target: beta Catenin (CATNB) (Catenin, beta (CATNB))

Binding Specificity: AA 571-781

Reactivity: Human, Mouse

Host: Mouse

Clonality: Monoclonal

Conjugate: This beta Catenin antibody is conjugated to Alexa Fluor 488

Application: Immunofluorescence (IF), BioImaging (BI), Intracellular Staining (ICS)

Brand: BD Pharmingen™

Purification: The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Immunogen: Mouse beta-Catenin aa. 571-781

Clone: 14-Beta

Isotype: IgG1

Application Details

Sample Volume: 5 μL

Protocol: 1. Seed ~10,000 cells in appropriate culture medium in a BD Falcon™ 96-well Imaging Plate (Cat. No. 353219) and culture overnight.
2. Remove the culture medium from the wells and fix the cells by adding 100 µl of fresh 3.7% Formaldehyde in PBS or BD Cytofix™ Fixation Buffer (Cat. No. 554655) to each well and incubate for 10 minutes at room temperature (RT).
3. Remove the fixative from the wells, and permeabilize the cells using either cold methanol or Triton™ X-100.
a. Add 100 myl of -20°C 90% methanol or -20°C BD Phosflow™ Perm Buffer III (Cat. No. 558050) to each well and incubate (5 min, RT). OR
b. Add 100 µl of 0.1% Triton™ X-100 (Sigma-Aldrich Cat. No. X-100) to each well and incubate (5 min, RT).
4. Remove the permeabilizer from the wells, and wash wells twice with 100 myl of 1x PBS.
5. Remove the PBS and block the cells by adding 100 myl of blocking buffer, eg, BD Pharmingen™ Stain Buffer (FBS) (Cat. No. 554656) or 3% FBS in 1x PBS to each well and incubate (30 min, RT).
6. Meanwhile, put 5 ul (1 test size) Bioimaging Certified fluorescent antibodies into 45 ul of Stain Buffer, protect from light.
7. Remove the blocking buffer and add 50 myl of prepared fluorescent antibody per well and incubate (60 min, RT), protect from light.
8. Remove the fluorescent antibody and wash the wells three times with 100 myl of 1x PBS.
9. Remove the PBS and counterstain the nuclei by adding 100 µl of a 2 µg/ml solution of Hoechst 33342 (Cat. No. 561908) in 1x PBS to each well at least 15 minutes before imaging.
10. View and analyze the cells using an appropriate imaging instrument.

Restrictions: For Research Use only

Handling

Format: Liquid

Buffer: Aqueous buffered solution containing BSA, protein stabilizer, and ≤0.09 % sodium azide.

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: 4 °C

Storage Comment: Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The antibody was conjugated to Alexa Fluor® 488 under optimum conditions, and unreacted Alexa Fluor® 488 was removed.

References for anti-beta Catenin antibody (Alexa Fluor 488) (ABIN1176964)

Lee, DAmour, Papkoff: "A yeast model system for functional analysis of beta-catenin signaling." in: The Journal of cell biology, Vol. 158, Issue 6, pp. 1067-78, (2002) (PubMed).

Persad, Troussard, McPhee, Mulholland, Dedhar: "Tumor suppressor PTEN inhibits nuclear accumulation of beta-catenin and T cell/lymphoid enhancer factor 1-mediated transcriptional activation." in: The Journal of cell biology, Vol. 153, Issue 6, pp. 1161-74, (2001) (PubMed).

Tateishi, Omata, Tanaka, Chiba: "The NEDD8 system is essential for cell cycle progression and morphogenetic pathway in mice." in: The Journal of cell biology, Vol. 155, Issue 4, pp. 571-9, (2001) (PubMed).

Ozawa, Ringwald, Kemler: "Uvomorulin-catenin complex formation is regulated by a specific domain in the cytoplasmic region of the cell adhesion molecule." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 87, Issue 11, pp. 4246-50, (1990) (PubMed).

Target Details for beta Catenin

Target: beta Catenin (CATNB) (Catenin, beta (CATNB))

Alternative Name: beta-Catenin (CATNB Products)

Synonyms: Bfc antibody, Catnb antibody, Mesc antibody, CTNNB antibody, MRD19 antibody, armadillo antibody, ctnnb antibody, id:ibd2058 antibody, wu:fb73e10 antibody, wu:fi81c06 antibody, wu:fk25h01 antibody, beta-catenin antibody, ctnnb1 antibody, CHBCAT antibody, catenin (cadherin associated protein), beta 1 antibody, catenin beta 1 antibody, catenin (cadherin-associated protein), beta 1 antibody, catenin beta 1 L homeolog antibody, Ctnnb1 antibody, CTNNB1 antibody, ctnnb1 antibody, ctnnb1.L antibody

Background: The 14/Beta-Catenin monoclonal antibody specifically binds to Beta-Catenin (beta-Catenin). beta-Catenin is a 92 kDa protein that binds to the cytoplasmic tail of E-Cadherin. The cadherins, transmembrane adhesion molecules, are found with catenins at adherens junctions (zonula adherens). Deletions in the cytoplasmic domain of E-Cadherin which eliminate catenin binding also result in a loss of cell adhesion, indicating that this binding is essential for E-Cadherin function. Although the alpha- and beta-Catenins have been cloned, very little is known about their biochemical roles. However a link between beta-Catenin and colon cancer has been described. beta-Catenin was found to co-immunoprecipitate with the APC tumor suppressor protein in human colorectal tumor cell lines, as well as in human kidney 293 cells. E-Cadherin, however, was not detectable in these complexes. Thus the APC-Catenin complex may be affecting the transmission of contact inhibition signals and/or the regulation of cell adhesion.
Synonyms: CTNNB1, CTNB1, Catenin beta-1, catenin (cadherin-associated protein) beta 1

Pathways: Peptide Hormone Metabolism