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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [2]
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- Product number
- MA5-17236 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Anti-Cathepsin D Monoclonal Antibody (AF4G5)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- Description
- A suggested positive control for this product is SK-BR3 cells.
- Reactivity
- Human, Mouse, Rat
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- AF4G5
- Vial size
- 100 µL
- Concentration
- 1 mg/mL
- Storage
- -20°C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Cathepsin D using anti-Cathepsin D monoclonal antibody (Product # MA5-17236) on (Lane 1) MCF7 Cell lysate, (Lane 2) HepG2 Cell lysate and SK-BR3 Cell lysate.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of Hep G2 (Lane 1), HeLa (Lane 2), MCF7 (Lane 3), 3T3-L1 (Lane 4), A549 (Lane 5) and MDA-MB-231 (Lane 6). The blots were probed with Cathepsin D Mouse monoclonal Antibody (Product # MA5-17236, 0.5 µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). A 52 kDa band corresponding to Cathepsin D was observed across the cell lines tested. Apart from desired band a non-specific band was observed in HeLa cell line. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX), 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 iBlot® 2 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 SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).
