AF523

antibody from R&D Systems

Targeting: IL1RL1 DER4, FIT-1, IL33R, ST2, ST2L, ST2V, T1

Western blot (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Blocking/Neutralizing (Supportive data in Antibodypedia)

Antibody Data

Product number

AF523

Provider

R&D Systems

Product name

Human ST2/IL-33R Antibody

Antibody type

Polyclonal

Description

Antigen Affinity-purified. Detects human ST2/IL-33 R in direct ELISAs and Western blots. In direct ELISAs, less than 10% cross-reactivity with recombinant mouse ST2/IL-33 R is observed.

Reactivity

Human

Host

Goat

Conjugate

Unconjugated

Antigen sequence

BAA02233


Isotype

IgG

Vial size

100 ug

Concentration

LYOPH

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.

References

Submitted references

IL-33 stimulates CXCL8/IL-8 secretion in goblet cells but not normally differentiated airway cells.

Tanabe T, Shimokawaji T, Kanoh S, Rubin BK
Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology 2014 Apr;44(4):540-52

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Detection of soluble ST2 in human follicular fluid and luteinized granulosa cells.

Southcombe JH, Lédée N, Perrier d'Hauterive S, Turner K, Child T, Snider JV, Redman CW, Sargent IL, Granne I
PloS one 2013;8(9):e74385

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ST2 and IL-33 in pregnancy and pre-eclampsia.

Granne I, Southcombe JH, Snider JV, Tannetta DS, Child T, Redman CW, Sargent IL
PloS one 2011;6(9):e24463

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Measurement of interleukin-33 (IL-33) and IL-33 receptors (sST2 and ST2L) in patients with rheumatoid arthritis.

Hong YS, Moon SJ, Joo YB, Jeon CH, Cho ML, Ju JH, Oh HJ, Heo YJ, Park SH, Kim HY, Min JK
Journal of Korean medical science 2011 Sep;26(9):1132-9

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Expression of interleukin 1-like cytokine interleukin 33 and its receptor complex (ST2L and IL1RAcP) in human pancreatic myofibroblasts.

Nishida A, Andoh A, Imaeda H, Inatomi O, Shiomi H, Fujiyama Y
Gut 2010 Apr;59(4):531-41

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A novel IL-1 family cytokine, IL-33, potently activates human eosinophils.

Cherry WB, Yoon J, Bartemes KR, Iijima K, Kita H
The Journal of allergy and clinical immunology 2008 Jun;121(6):1484-90

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Gene induction during differentiation of human pulmonary type II cells in vitro.

Wade KC, Guttentag SH, Gonzales LW, Maschhoff KL, Gonzales J, Kolla V, Singhal S, Ballard PL
American journal of respiratory cell and molecular biology 2006 Jun;34(6):727-37

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IL-33, an interleukin-1-like cytokine that signals via the IL-1 receptor-related protein ST2 and induces T helper type 2-associated cytokines.

Schmitz J, Owyang A, Oldham E, Song Y, Murphy E, McClanahan TK, Zurawski G, Moshrefi M, Qin J, Li X, Gorman DM, Bazan JF, Kastelein RA
Immunity 2005 Nov;23(5):479-90

Western blot

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

Detection of Human ST2/IL-33 R by Western Blot. Western blot shows lysates of human kidney tissue and human lung tissue. PVDF membrane was probed with 0.2 µg/mL of Goat Anti-Human ST2/IL-33 R Antigen Affinity-purified Polyclonal Antibody (Catalog # AF523) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for ST2/IL-33 R at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

Detection of Human ST2/IL-33 R by Simple Western^TM. Simple Western lane view shows lysates of human kidney tissue and human lung tissue, loaded at 0.2 mg/mL. A specific band was detected for ST2/IL-33 R at approximately 60 kDa (as indicated) using 2 µg/mL of Goat Anti-Human ST2/IL-33 R Antigen Affinity-purified Polyclonal Antibody (Catalog # AF523) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Immunohistochemistry

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

ST2/IL-33 R in Human Liver. ST2/IL-33 R was detected in immersion fixed paraffin-embedded sections of human liver using Goat Anti-Human ST2/IL-33 R Antigen Affinity-purified Polyclonal Antibody (Catalog # AF523) at 1.7 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cell membranes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Flow cytometry

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

Detection of ST2/IL-33 R in KG-1a Human Cell Line by Flow Cytometry. KG-1a human acute myelogenous leukemia cell line was stained with Goat Anti-Human ST2/IL-33 R Antigen Affinity-purified Polyclonal Antibody (Catalog # AF523, filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram), followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108).

Blocking/Neutralizing

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

IFN-gamma Secretion Induced by IL-33 and Neutralization by Human ST2/IL-33 R Antibody. Recombinant Human IL-33 induces IFN-gamma secretion in Human Peripheral Blood Mononuclear cells (PBMC) in the presence of 0.25 ng/mL Recombinant Human IL-12 (Catalog # 219-IL) in a dose-dependent manner (orange line), as measured by the Human IFN-gamma Q-kit (Catalog # DIF50). Under these conditions, IFN-gamma secretion elicited by IL-33 is neutralized (green line) by increasing concentrations of Goat Anti-Human ST2/IL-33 R Antigen Affinity-purified Polyclonal Antibody (Catalog # AF523). The ND₅₀ is typically 0.1-0.6 μg/mL.