Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [4]
- Immunocytochemistry [5]
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- Product number
- PA5-20562 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- NUP107 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- A suggested positive control is A549 cell lysate. PA5-20562 can be used with blocking peptide PEP-0682.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µg
- Concentration
- 1 mg/mL
- Storage
- Maintain refrigerated at 2-8°C for up to 3 months. For long term storage store at -20°C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of A549 cell lysate using a NUP107 polyclonal antibody (Product # PA5-20562) at (A) 1 and (B) 2 µg/mL.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of NUP107 in A549 cell lysate with NUP107 Polyclonal Antibody (Product # PA5-20562) at (A) 1 and (B) 2 µg/mL.
- 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 # PA5-20562, 2 µg/mL) 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. Few uncharacterized bands were also observed at ~70kDa and ~35kDa.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-NUP107 Polyclonal Antibody (Product # PA5-20562) and 107 kDa, 78 kDa bands corresponding to NUP107 was observed across all the cell lines and tissues tested along with an uncharacterized band at ~ 30 kDa (*). Modified whole cell extracts (1% SDS) (30 µg lysate) of HeLa (Lane 1), A549 (Lane 2), HEK-293 (Lane 3), MCF7 (Lane 4), PC-3 (Lane 5), NIH/3T3 (Lane 6), tissue extracts of Mouse Brain (Lane 7) and Mouse Kidney (Lane 8)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 (2 µg/mL) 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
- Immunocytochemistry staining of A549 cells using a NUP107 polyclonal antibody (Product # PA5-20562) at a 10 µg/mL dilution.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of A549 cells using a NUP107 polyclonal antibody (Product # PA5-20562) at a 10 µg/mL dilution.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry of NUP107 in A549 cells with NUP107 Polyclonal Antibody (Product # PA5-20562) at 10 µg/mL.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence of NUP107 in A549 cells with NUP107 Polyclonal Antibody (Product # PA5-20562) at 10 µg/mL.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of NUP107 was performed using 70% confluent log phase PC-3 cells. The cells were fixed with 4% Paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 10 minutes at room temperature. The cells were labeled with NUP107 Polyclonal Antibody (Product # PA5-20562) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790, 1:2000 dilution) for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear membrane localization for NUP107 in PC-3. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.