Antibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [3]
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Validation data
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- Product number
- PA5-17531 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- IRAK4 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- It is not recommended to aliquot this antibody.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 300 µg/mL
- Storage
- -20°C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of IRAK4 in extracts from RAW 264.7 and H-4-II-E cell lines using IRAK4 polyclonal antibody (Product # PA5-17531).
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Knockdown of IRAK4 was achieved by transfecting THP-1 cells with IRAK4 specific siRNAs (Silencer® select Product # s535019, s535020). Western blot analysis (Fig. a) was performed using Whole Cell Extract from the IRAK4 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with IRAK4 Polyclonal Antibody (Product # PA5-17531, 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 IRAK4, however an uncharacterized (*) band was observed at ~60 kDa.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-IRAK4 Polyclonal Antibody (Product # PA5-17531) and a 52 kDa band corresponding to IRAK4 was observed across cell lines tested. Whole Cell Extract (40 µg lysate) of THP-1 (Lane 1), THP-1 treated with PMA (25 nM for 48 hours) and LPS (100 ng/ml for 24 hours) (Lane 2), THP-1 (Lane 3), THP-1 subjected to M0 macrophage differentiation and treated with LPS (100ng/ml for 12 hours) (Lane 4) 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). THP-1 differentiated macrophage upon LPS treatment are reported to show down regulation in IRAK4 levels (doi:10.1016/j.cellsig.2008.10.009).