Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [4]
- Immunocytochemistry [1]
- Immunohistochemistry [2]
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Validation data
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- Product number
- PA5-27651 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- DPF2 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: DPF2 tansfected 293T cell.
- Concentration
- 0.67 mg/mL
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of DPF2 expression in transfected 293T cell line by DPF2 polyclonal antibody. Lane A: Non-transfected lysate. Lane B: DPF2 transfected lysate. 12% SDS PAGE. DPF2 Polyclonal Antibody (Product # PA5-27651) diluted at 1:500.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-DPF2 Polyclonal Antibody (Product # PA5-27651) and a 45 kDa band corresponding to Zinc finger protein ubi-d4 was observed along with uncharacterized bands (*) across cell lines and tissues tested. Nuclear enriched extracts (30 µg lysate) of A-431 (Lane 1), A549 (Lane 2), HEL 92.1.7 (Lane 3), K-562 (Lane 4) and NIH/3T3 (Lane 5). Tissue extracts of Mouse Thymus (Lane 6) and Rat Thymus (Lane 7) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). 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).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of DPF2 expression in transfected 293T cell line by DPF2 polyclonal antibody. Lane A: Non-transfected lysate. Lane B: DPF2 transfected lysate. 12% SDS PAGE. DPF2 Polyclonal Antibody (Product # PA5-27651) diluted at 1:500.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of Zinc finger protein ubi-d4 was achieved by transfecting K-562 with Zinc finger protein ubi-d4 specific siRNAs (Silencer® select Product # S11929, S11930). Western blot analysis (Fig. a) was performed using Nuclear enriched extracts from the Zinc finger protein ubi-d4 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with DPF2 Polyclonal Antibody (Product # PA5-27651, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 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 Zinc finger protein ubi-d4.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Zinc finger protein ubi-d4 was performed using 70% confluent log phase Hep G2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with DPF2 Polyclonal Antibody (Product # PA5-27651) at 5 µg/mL 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 Hoechst 33342 (Product # H1399). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing Nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 40X magnification in CellInsight CX7 LZR High-Content Screening (HCS) Platform (Product # CX7A1110LZR) and externally deconvoluted (D.Sage et al. / Methods 115 (2017) 28–41).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry (Paraffin) analysis of DPF2 was performed in paraffin-embedded mouse brain tissue using DPF2 Polyclonal Antibody (Product # PA5-27651) at a dilution of 1:500.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of paraffin-embedded H661 xenograft, using DPF2 (Product # PA5-27651) antibody at 1:250 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.