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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [2]
- Immunocytochemistry [1]
- Other assay [1]
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- Product number
- PA5-47359 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- IL1R2 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- Description
- In direct ELISAs, less than 1% cross-reactivity with recombinant human (rh) IL-1 alpha, rhIL-1 RI, recombinant mouse (rm) IL-1 alpha, rhIL-1 beta, rmIL-1 beta, recombinant rat IL-1 beta, rhIL-1ra, rmIL-1ra, and recombinant rhesus monkey IL-1ra is observed.
- Concentration
- 0.2 mg/mL
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of Interleukin-1 receptor type 2 was achieved by transfecting HT-29 with Interleukin-1 receptor type 2 specific siRNAs (Silencer® select Product # s15407, s15408 ). Western Blot analysis (Fig. a) was performed using Membrane enriched extracts from the Interleukin-1 receptor type 2 knockdown cells (lane 3), non-targeting siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with IL1R2 Polyclonal Antibody (Product # PA5-47359, 0.3 µg/mL) and Rabbit anti-Goat IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27014, 1:6000 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 Interleukin-1 receptor type 2.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot was performed using Anti-IL1R2 Polyclonal Antibody (Product # PA5-47359) and a 68 kDa band (membrane bound isoform) corresponding to Interleukin-1 receptor type 2 was observed across cell lines tested. Membrane enriched extracts (30 µg lysate) of HeLa (Lane 1), HT-29 (Lane 2), COLO 205 (Lane 3), HCT 116 (Lane 4) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0301BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (0.3 µg/mL) and detected by chemiluminescence with Rabbit anti-Goat IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27014, 1:6000 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
- Immunocytochemistry analysis of IL1R2 in immersion fixed human peripheral blood mononuclear cells (PBMCs). Samples were incubated in IL1R2 polyclonal antibody (Product # PA5-47359) using a dilution of 15 µg/mL for 3 hours at room temperature. Cells were stained (red) and counterstained (green).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Neutralization of IL1R2 in D10.G4.1 mouse helper T cell line. Samples were incubated in IL1R2 polyclonal antibody (Product # PA5-47359). Recombinant Human IL‚1 RII inhibits Recombinant Human IL‚1β/IL‚1F2 induced proliferation in the D10.G4.1 mouse helper T cell line in a dose-dependent manner (orange line). Inhibition of Recombinant Human IL‚1β/IL‚1F2 (50 pg/mL) activity elicited by Recombinant Human IL‚1 RII (2 µg/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human IL‚1 RII Antigen Affinity-purified Polyclonal Antibody. The ND50 is typically 15‚30 µg/mL.
