Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [1]
- Immunocytochemistry [1]
Submit
Validation data
Reference
Comment
Report error
- Product number
- 711404 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Phospho-beta Catenin (Ser33, Ser37) Recombinant Polyclonal Antibody (4HCLC)
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- 4HCLC
- Vial size
- 100 µg
- Concentration
- 0.5 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
No comments: Submit comment
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of SW480 (Lane 1) and SW480 treated with MG132 (25uM for 4 hrs) (Lane 2). The blots were probed with Anti-β-Catenin p (S33/S37) Recombinant Rabbit Polyclonal Antibody (Product # 711404, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 85 kDa band corresponding to β-Catenin p (S33/S37) was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis Systm (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- For immunofluorescence analysis, SW480 cells were fixed and permeabilized for detection of endogenous beta-Catenin pS33 pS37 using Anti-Beta-Catenin pS33 pS37 Recombinant Rabbit Polyclonal Antibody (Product # 711404, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Nuclei (blue) were stained using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938), and Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300) was used for cytoskeletal F-actin (red) staining. Detection and localization of beta-Catenin pS33 pS37 (green) in the cytoplasm can be clearly observed in cells treated with the proteasome inhibitor MG-132 (25 uM, 4 h) as compared to untreated cells. Antibody specificity was demonstrated by competition with the beta-Catenin pS33 pS37 peptide, which results in loss of signal. No competition was observed with the non-phospho peptide. The images were captured at 60X magnification.