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
- PA5-19504 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- IRF5 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- Not suitable for use with Immunohistochemistry (Paraffin). This antibody is predicted to react with cow based on sequence homology.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µg
- Concentration
- 0.9 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 Mouse Liver Tissue Lysate using Product # PA5-19504, IRF5 primary antibody at a dilution of 1 µg/mL (lane 1). Staining of Rat Liver Tissue Lysate at a dilution of 1 µg/mL (lane 2). Blot treated with a secondary IR Dye680-conjugated Goat polyclonal anti-Rabbit antibody was used at a dilution of 1:10000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-IRF5 Polyclonal Antibody, (Product # PA5-19504) and a 57 kDa band corresponding to IRF5 along with uncharacterized bands (*) at ~33 kDa and 20 kDa were observed in the cell lines tested with higher expression in THP-1, HEL 92.1.7 compared to Jurkat and K-562. Modified whole cell extracts (1% SDS) (40 µg lysate) of THP-1 (Lane 1), Jurkat (Lane 2), K-562 (Lane 3), and HEL 92.1.7 (Lane 4) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris 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 µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant 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 IRF5 was performed using 70% confluent log phase THP-1 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 1 hour at room temperature. The cells were labeled with IRF5 Polyclonal Antibody (Product # PA5-19504) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790) at a dilution of 1:2000 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 cytoplasmic and nuclear localization. Panel e represents Jurkat cells having no expression of IRF5. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.