Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunohistochemistry [2]
- Flow cytometry [1]
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Validation data
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- Product number
- PA1-9057 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- IRAK4 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- This antibody is tested in Peptide ELISA: antibody detection limit dilution 32,000.
- Reactivity
- Human
- Host
- Goat
- Isotype
- IgG
- Vial size
- 100 µg
- Concentration
- 0.5 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot detection of IRAK4 in hela lysate using Product # PA1-9057.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of IRAK4 was achieved by transfecting THP-1 with IRAK4 specific siRNAs (Silencer® select Product # s535019, s535020). Western blot analysis (Fig. a) was performed using whole cell extracts from the IRAK4 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with IRAK4 Polyclonal Antibody (Product # PA1-9057, 1:500 dilution) and Rabbit anti-Goat IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP conjugate (Product # A27014, 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.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-IRAK4 Polyclonal Antibody (Product # PA1-9057) and a 52 kDa band corresponding to IRAK4 was observed across the cell lines tested. Whole cell extracts (40 µg lysate) of THP-1 (Lane 1), HeLa (Lane 2), RAW 264.7 (Lane 3) and Jurkat (Lane 4) 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 (1:500 dilution) and detected by chemiluminescence with Rabbit anti-Goat IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27014, 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
- Immunohistochemical analysis of IRAK4 in paraffin embedded human heart using a IRAK4 polyclonal antibody (Product #PA1-9057) at a concentration of 3.75 µg/mL. Steamed antigen retrieval was performed with pH 6 buffered citrate. Samples were then stained with alkaline phosphatase.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of IRAK4 in paraffin embedded human heart using a IRAK4 polyclonal antibody (Product #PA1-9057) at a concentration of 3.75 µg/mL. Steamed antigen retrieval was performed with pH 6 buffered citrate. Samples were then stained with alkaline phosphatase.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometric analysis of IRAK4 in HeLa cells using a polyclonal antibody (Product #PA1-9057). HeLa cells (blue line) were paraformaldehyde fixed and permeabilized with 0.5% Triton. The primary antibody was incubated for one hour (10 µg/mL) followed by an Alexa Fluor 488 secondary antibody (1 µg/mL). IgG control: Unimmunized goat IgG (black line) followed by an Alexa Fluor 488 secondary antibody.