IL-1 RII Antibody (32437) [Unconjugated]

Recombinant Human IL-1 RII (263-2R) inhibits Recombinant Human IL-1 beta / IL-1F2 (201-LB) induced proliferation in the D10.G4.1 mouse helper T cell line in a dose-dependent manner (orange line). Inhibition of Recombinant Human IL-1 beta /IL-1F2 (50 pg/mL) activity elicited by Recombinant Human IL-1 RII (2 µg/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human IL-1 RII Monoclonal Antibody (Catalog # MAB263). The ND50 is typically 2-6 µg/mL.

IL‑1 RII was detected in immersion fixed HDLM‑2 human Hodgkin’s lymphoma cells (positive) and absent in K562 human chronic myelogenous leukemia cells (negative) using Mouse Anti-Human IL‑1 RII Monoclonal Antibody (Catalog # MAB263) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cell surface. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

HDLM-2 cells were stained with Mouse Anti-Human IL‑1 RII Monoclonal Antibody (Catalog # MAB263, filled histogram) or isotype control antibody (Catalog # MAB003, open histogram) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). View our protocol for Staining Membrane-associated Proteins.

Isolation of SP cells from human first trimester vCTB.(A) Immunocytochemistry images of CGB, CK7, CSH1, HLA−G, CD9 and HLA1 expression in primary vCTB cells isolated by Percoll gradient centrifugation. Each protein expression is shown as red signals (scale bars, 50 µm). The isolated cells were plated on chamber slides with HBM and stained the next day. (B) The SP cells represent 0.12±0.28% (n = 22) of the total primary vCTB. Co-incubation of the cells with verapamil erased the SP cell fraction. (C) The expression of CD105, CD146 and VIM in SP cells from primary vCTB cells was analyzed by immunocytochemistry. All images show counterstaining with Hoechst 33852. Each protein expression is shown as red signals (scale bars, 50 µm). (D) Immunocytochemistry images of IL7R and IL1R2 expression in SP and NSP cells counterstained with Hoechst 33852. IL7R and IL1R2 expressions are shown as red signals (scale bars, 50 µm). (E) The expression of CGB, CSH1 HLA−G, CK7, PHLDA2 and EOMES in IL7R/IL1R2 double-positive cells was assessed by immunocytochemistry. These expressions are shown as green signals (scale bars, 50 µm). (F) Induction of differentiation in IL7R/IL1R2 positive cells was determined by IL7R, IL1R2, CSH1 and CGB expression. IL7R/IL1R2 positive cells from primary human vCTB were analyzed after being cultured in HBM for 2 weeks. IL7R and IL1R2 expressions are shown as red signals; CSH1 and CGB expressions are shown as green signals (scale bars, 50 µm). The cell marked with an arrow show positive for CGB and negative for IL7R expression. In contrast, the cell marked with an arrowhead was positive for IL7R and negative for CGB expression. The outlines of the cells are indicated by broken lines. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/21760941), licensed under a CC-BY license. Not internally tested by R&D Systems.

IL7R and IL1R2 were abundantly expressed in HTR-8/SVneo-SP cells.(A) IL7R and IL1R2 mRNA expressions between HTR-8/SVneo-SP and -NSP cells were compared by real-time RT-PCR. The mRNA levels were normalized to those of GAPDH and error bars show standard deviations (n = 3). Asterisk indicates statistically significant (p<0.01). (B) Western blot analysis of IL7R, IL1R2 and CXCR7 proteins in HTR-8/SVneo-SP and -NSP cells. SP cells exclusively expressed IL7R and IL1R2, while NSP exclusively expressed CXCR7. (C) The protein expression of IL7R, IL1R2 or CXCR7 was analyzed by immunocytochemistry. All images counterstained with Hoechst 33852. Each protein expression is shown as green signals (scale bars, 50 µm). (D) IL7R and IL1R2 expressions in HTR-8/SVneo-SP cells were analyzed by immunocytochemistry. All images were counterstained with Hoechst 33852. IL7R and IL1R2 expressions are shown as red and green signals respectively (scale bars, 50 µm). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/21760941), licensed under a CC-BY license. Not internally tested by R&D Systems.

SP and IL1R2/IL7R double-positive cells differentiated into multiple trophoblast cell lineages.(A) Immunocytochemistry images of IL7R, IL1R2, CK7, CSH1, CGB and HLA−G expression in SP cells sorted from HTR-8/SVneo parent cells. Each protein expression is shown as green signals (scale bars, 50 µm). The cells were assayed after culture for 2 weeks in HSM or HBM. (B) Flow cytometry results of IL1R2-positive cells after being sorted by MACS. SP cells were obtained from IL1R2-positive cells after culture for 2 weeks (4.78±1.45% from 3 independent experiments). (C) Induction of trophoblast differentiation of IL7R/IL1R2 double-positive HTR-8/SVneo cells confirmed by differentiation marker, CSH1 and CGB expression. IL7R and IL1R2 expressions are shown as red signals; CSH1 and CGB expressions are shown as green signals (scale bars, 50 µm). The cells marked with an arrow show positive for CSH1 or CGB and negative for IL1R2 or IL7R expression. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/21760941), licensed under a CC-BY license. Not internally tested by R&D Systems.

IL7R and IL1R2 were abundantly expressed in HTR-8/SVneo-SP cells.(A) IL7R and IL1R2 mRNA expressions between HTR-8/SVneo-SP and -NSP cells were compared by real-time RT-PCR. The mRNA levels were normalized to those of GAPDH and error bars show standard deviations (n = 3). Asterisk indicates statistically significant (p<0.01). (B) Western blot analysis of IL7R, IL1R2 and CXCR7 proteins in HTR-8/SVneo-SP and -NSP cells. SP cells exclusively expressed IL7R and IL1R2, while NSP exclusively expressed CXCR7. (C) The protein expression of IL7R, IL1R2 or CXCR7 was analyzed by immunocytochemistry. All images counterstained with Hoechst 33852. Each protein expression is shown as green signals (scale bars, 50 µm). (D) IL7R and IL1R2 expressions in HTR-8/SVneo-SP cells were analyzed by immunocytochemistry. All images were counterstained with Hoechst 33852. IL7R and IL1R2 expressions are shown as red and green signals respectively (scale bars, 50 µm). Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/21760941), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, B/N, ICC/IF
• Clone: 32437
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

IL-1 RII Antibody (32437) [Unconjugated] Summary

• Immunogen: S. frugiperda insect ovarian cell line Sf 21-derived recombinant human IL‑1 RII
  Phe14-Glu343 (Glu297Gly)
  Accession # P27930
• Specificity: Detects human IL-1 RII in direct ELISAs and Western blots. In direct ELISAs, no cross-reactivity with recombinant human (rh) IL-1 RI, rhIL‑18 R, and recombinant mouse IL-18 R is observed.
• Source: N/A
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Mouse
• Gene: IL1R2
• Purity Statement: Protein A or G purified from ascites
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 8-25 ug/mL
  • Neutralization 2-6 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
  *Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for IL-1 RII Antibody (32437) [Unconjugated]

• beta
• CD121b antigen
• CD121b
• IL-1 R beta
• IL-1 RII
• IL1R2
• IL1RBCD121 antigen-like family member B
• IL-1R-beta
• IL1RII
• IL-1RII
• IL-1RT2
• IL-1RT-2
• interleukin 1 receptor, type II
• Interleukin-1 receptor beta
• MGC47725

Background

Two distinct types of receptors that bind the pleiotropic cytokines IL-1 alpha and IL-1 beta have been described. The IL-1 receptor type I is an 80 kDa transmembrane protein that is expressed predominantly by T cells, fibroblasts and endothelial cells. IL-1 receptor type II is a 68 kDa transmembrane protein found on B lymphocytes, neutrophils, monocytes, large granular leukocytes and endothelial cells. Both receptors are members of the immunoglobulin superfamily and show approximately 28% sequence similarity in their extracellular domains. The two receptor types do not heterodimerize in a receptor complex.

An IL-1 receptor accessory protein that can heterodimerize with the type I receptor in the presence of IL-1 alpha or IL-1 beta but not IL-1ra, was identified (1). This type I receptor complex appears to mediate all the known IL-1 biological responses. The receptor type II has a short cytoplasmic domain and does not transduce IL-1 signals. In addition to the membrane-bound form of IL-1 RII, a naturally-occurring soluble form of IL-1 RII has been described. It has been suggested that the type II receptor, either as the membrane-bound or as the soluble form, serves as a decoy for IL-1 and inhibits IL-1 action by blocking the binding of IL-1 to the signaling type I receptor complex.

1. Greenfeder, S. et al. (1995) J. Biol. Chem. 270:13757.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for IL-1 RII Antibody (MAB263)(3)

Publications using MAB263

• Dong Hyun Kim, Hee Young Kim, Sunjung Cho, Su-Jin Yoo, Won-Ju Kim, Hye Ran Yeon et al. Induction of the IL-1RII decoy receptor by NFAT/FOXP3 blocks IL-1 beta -dependent response of Th17 cells eLife 2021-01-28 [PMID: 33507149]
• Takao T, Asanoma K, Kato K Isolation and characterization of human trophoblast side-population (SP) cells in primary villous cytotrophoblasts and HTR-8/SVneo cell line. PLoS ONE, 2011-07-07;6(7):e21990. 2011-07-07 [PMID: 21760941] (Cell Selection, ICC, Western Blot, Human)
• Grundtman C, Salomonsson S, Dorph C, Bruton J, Andersson U, Lundberg IE Immunolocalization of interleukin-1 receptors in the sarcolemma and nuclei of skeletal muscle in patients with idiopathic inflammatory myopathies. Arthritis Rheum., 2007-02-01;56(2):674-87. 2007-02-01 [PMID: 17265504] (ICC, IHC-Fr, Human)

Bioinformatics

• Gene Symbol: IL1R2
• Uniprot:
  • P27930()