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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
- PA5-17767 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Ezrin Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- It is not recommended to aliquot this antibody. This antibody is not cross-reactive with related proteins such as merlin or band 4.
- Reactivity
- Human, Mouse, Rat, Bovine, Canine, Xenopus
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Storage
- -20°C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of Ezrin was achieved by transfecting HCT 116 with Ezrin specific siRNAs (Silencer® select Product # s14796, s14795). Western blot analysis (Fig. a) was performed using Membrane enriched extracts from the Ezrin knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with Ezrin Polyclonal Antibody (Product # PA5-17767, 1:1000) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000). 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 Ezrin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-Ezrin Polyclonal Antibody(Product # PA5-17767) and a ~70kDa band corresponding to Ezrin was observed across cell lines tested . Membrane enriched extracts (30 µg lysate) of HeLa (Lane 1), HCT 116 (Lane 2), A-431 (Lane 3), HaCaT (Lane 4), NTERA-2 cl.D1 (Lane 5), HEK-293 (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Increased expression was observed in HeLa, HCT 116, A-431 and HaCaT compared to in NTERA and HEK-293. A non specific band was observed around 15kDa in HeLa, HaCaT and NTERA cell lines.
