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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
- LF-MA0074 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Anti-14-3-3 beta Monoclonal Antibody (60C10)
- Antibody type
- Monoclonal
- Antigen
- Recombinant full-length protein
- Description
- A suggested positive control for this product is HeLa cells.
- Reactivity
- Human, Mouse, Rat
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 60C10
- Vial size
- 100 µL
- Concentration
- 1 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of 14-3-3 beta protein (YWHAB) was achieved by transfecting HeLa cells with YWHAB specific siRNAs (Silencer® select Product # s14962). Western blot analysis (Fig. a) was performed using membrane enriched cell extracts from the YWHAB knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed using YWHAB Monoclonal Antibody (Product # LF-MA0074, 1:1000 dilution) and Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to YWHAB.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-14-3-3 beta protein Monoclonal Antibody (Product # LF-MA0074) on membrane enriched cell extracts (30 µg lysate) of A-431 (Lane 1), MCF7 (Lane 2), U-87 MG (Lane 3), HEK-293 (Lane 4), HeLa (Lane 5), Mouse brain (Lane 6), Mouse Skeletal muscle (Lane 7), Rat Brain (Lane 8), Rat Skeletal muscle (Lane 9) and 28 kDa band corresponding to 14-3-3 beta protein was observed except in Mouse and Rat Skeletal Muscle. Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
