Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [1]
- Chromatin Immunoprecipitation [1]
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Validation data
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- Product number
- PA5-76462 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- IFI16 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- Description
- The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen and the purity is > 95% (by SDS-PAGE).
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 1 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of IFI16. Samples were incubated with IFI16 polyclonal antibody (Product # PA5-76462).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-IFI16 Polyclonal Antibody (Product # PA5-76462) and bands at 82, 88 kDa corresponding to IFI16 was observed across the cell lines tested and the expression was increased or detected in HeLa and HUVEC upon IFN gamma and IFN beta treatments respectively. Modified whole cell extracts (1% SDS) (30ug lysate) of HeLa (Lane 1), HeLa treated with IFN gamma (15ng/ml for 16hr) (Lane 2), HUVEC (Lane 3), HUVEC treated with IFN beta (1000 IU/ml for 6hr) (Lane 4), Raji (Lane 5), Ramos (Lane 6), Jurkat (Lane 7) and Daudi (Lane 8) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris 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 (1:500 dilution) and detected by chemiluminescence Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (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
- Immunofluorescence analysis of IFI16 was performed using HeLa cells treated with IFN Gamma. 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 1 hour at room temperature. The cells were labeled with IFI16 Polyclonal Antibody (Product # PA5-76462) at 5 µg/mL in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the composite image showing increased cytoplasmic expression of IFI16 upon treatment with IFN gamma. Panel e represents control cells showing cytoplasmic staining. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Chromatin Immunoprecipitation (ChIP) assay of endogenous IFI16 protein using Anti-IFI16 Antibody: ChIP was performed using Anti-IFI16 Rabbit polyclonal Antibody (Product # PA5-76462, 5 µg) on sheared chromatin from poly (dA:dT) transfected U2OS cells using the MAGnify ChIP System kit (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR using primers binding to c-MYC promoter, IL6 intron1 and SAT2 satellite repeats. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.