PA5-47359

antibody from Invitrogen Antibodies

Targeting: IL1R2 CD121b, IL1RB

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Blocking/Neutralizing (Data presented on provider website)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-47359

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. Reconstitute at 0.2 mg/mL in sterile PBS. Endoxin level is

Reactivity

Human

Host

Goat

Isotype

IgG

Vial size

100 µg

Concentration

0.2 mg/mL

Storage

-20° C, Avoid Freeze/Thaw Cycles

Western blot

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.

Supportive validation

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).

Immunocytochemistry

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).

Other assay

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.