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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [3]
- Other assay [1]
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- Product number
- PA5-30774 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- NUP107 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: A549, HeLa, HepG2, HCT116.
- Concentration
- 0.44 mg/mL
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using NUP107 Polyclonal Antibody (Product # PA5-30774). Sample (30 µg of whole cell lysate). Lane A: Hela . 7.5% SDS PAGE. NUP107 Polyclonal Antibody (Product # PA5-30774) diluted at 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of NUP107 was achieved by transfecting HEK-293 with NUP107 specific siRNAs (Silencer® select Product # s32728, s32727). Western blot analysis (Fig. a) was performed using modified whole cell extracts from the NUP107 knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with NUP107 Polyclonal Antibody (Product # PA5-30774, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 0.25µg/ml, 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 NUP107.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-NUP107 Polyclonal Antibody (Product # PA5-30774) and a 107 kDa band corresponding to NUP107 was observed across all the cell lines tested along with an uncharacterized band (*) at ~ 50 kDa. Modified whole cell extracts (1% SDS) (30 µg lysate) of HEK-293 (Lane 1), MCF7 (Lane 2) and PC-3 (Lane 3) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). 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 chemiluminescence with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- NUP107 Polyclonal Antibody immunoprecipitates NUP107 protein in IP experiments. IP Sample: 293T whole cell lysate/extract A : 30 µg whole cell lysate/extract of NUP107 protein expressing 293T cells B : Control with 3 µg of pre-immune rabbit IgG C : Immunoprecipitation of NUP107 by 3 µg of NUP107 Polyclonal Antibody (Product # PA5-30774) 7.5% SDS-PAGE The immunoprecipitated NUP107 protein was detected by NUP107 Polyclonal Antibody (Product # PA5-30774) diluted at 1 : 500. Anti-rabbit IgG (HRP) was used as a secondary reagent.
