Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [1]
- Immunocytochemistry [1]
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Validation data
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- Product number
- 711366 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Phospho-c-Abl (Tyr393) Recombinant Polyclonal Antibody (19HCLC)
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- 19HCLC
- 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.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of K562 (Lane 1), Jurkat (Lane 2), KARPAS 299 (Lane 3), Raji (Lane 4), Ramos (Lane 5), A549 (Lane 6), MDA-MB-231 (Lane 7) and U-2 OS (Lane 8). The blots were probed with Anti-CAbl (pY393) Isoform A Recombinant Rabbit Polyclonal Antibody (Product # 711366, 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 123 kDa band corresponding to CAbl (pY393) Isoform A 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 System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with overnight wet transfer System. 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 K-562 cells were fixed and permeabilized for detection of endogenous CAbl pY393 Isoform A using Anti-CAbl pY393 Isoform A Recombinant Rabbit Polyclonal Antibody (Product # 711366, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of CAbl pY393 protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating cytoplasmic localization of CAbl pY393. Panel e) shows loss of signal by competition with the CAblpY393 peptide demonstrating antibody specificity, and panel f) demonstrates no competition with the non-phospho peptide. Panel g) shows untreated cells with no signal. The images were captured at 60X magnification.