Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [1]
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Validation data
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- Product number
- PA5-17149 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- TNFAIP3 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- It is not recommended to aliquot this antibody.
- Reactivity
- Human
- 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
- Western blot analysis of A20/TNFAIP3 in extracts from HeLa cells, untreated or treated with IL-1alpha (20 ng/mL for 2 hours) using A20/TNFAIP3 polyclonal antibody (Product # PA5-17149).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-TNFAIP3 Polyclonal Antibody (Product # PA5-17149) and a 89kDa band corresponding to TNFAIP3 was observed across all cell lines and tissue extracts tested and was upregulated upon IL-1 alpha, TNF-alpha and LPS treatments. A uncharacterized band (*) was also observed in some cell lines at ~32kDa. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), HeLa treated with IL-1 alpha (20ng/ml for 4H) (Lane 2), HeLa treated with TNF-alpha (20ng/ml for 4H) (Lane 3), THP-1 (Lane 4), RAW 264.7 (Lane 5), RAW 264.7 treated with LPS (1ug/ml for 6H) (Lane 6), Jurkat (Lane 7) and Mouse Spleen (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 (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 SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of TNFAIP3 was performed using 70% confluent log phase HeLa 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 TNFAIP3 Polyclonal Antibody (Product # PA5-17149) 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 ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing cytoplasmic and nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.