Q: Assay Diluent RD1-63 contains visible precipitate. Is this acceptable to use?

A: A small amount of precipitate is expected and acceptable in Assay Diluent RD1-63. Variability has been observed between vials and diluent lots. It may not fully resolubilize but this will not impact assay performance. It is not recommended to filter the precipitate.

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Human ST2/IL-33R Quantikine ELISA Kit

R&D Systems | Catalog # DST200

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Citations (77)

Product Documents

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Product Summary
Product Specifications
Sample Values
Precision
Recovery
Linearity
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices
Research Areas

Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Serum (10 µL), EDTA Plasma (10 µL), Heparin Plasma (10 µL)

Sensitivity

13.5 pg/mL

Assay Range

31.3-2000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)

View all ST2/IL-33R ELISA Kits »

Product Summary for Human ST2/IL-33R Quantikine ELISA Kit

The Quantikine Human ST2/IL-33 R Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human ST2 in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant human ST2 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human ST2 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring ST2.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Human

Specificity

Natural and recombinant human ST2 as well as free ST2 and IL-33 complexed ST2

Cross-reactivity

< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.

Interference

No significant interference observed with available related molecules.

Sample Values

Serum/Plasma - Samples from apparently healthy volunteers were evaluated for the presence of human ST2 in this assay. No medical histories were available for the donors used in this study.

Sample Type	Mean (ng/mL)	Range (ng/mL)	Standard Devitation (ng/mL)
Serum (n=35)	13.0	6.74-20.4	4.15
Heparin plasma (n=35)	12.3	5.72-19.8	4.10
EDTA plasma (n=35)	12.2	4.90-19.9	4.08

Cell Culture Supernates: BJAB human Burkitt's lymphoma cells (1 x 105 cells/mL) were seeded in DMEM supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. An aliquot of the cell culture supernate was removed, assayed for human ST2, and measured 126 pg/mL.

BUD-8 human fibroblast cells were seeded in MEM NEAA 90% supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate until confluent. An aliquot of the cell culture supernate was removed, assayed for human ST2, and measured 273 pg/mL.

U-87 MG human glioblastoma/astrocytoma cells (2.1 x 105 cells/mL) were seeded in MEM supplemented with 10% fetal bovine serum, 1% sodium pyruvate, and 2 mM L-glutamine until confluent. An aliquot of the cell culture supernate was removed, assayed for human ST2, and measured 63.9 pg/mL.

HUVEC human umbilical vein endothelial cells (1 x 104 cells/mL) were seeded in EGM-2 media until confluent. An aliquot of the cell culture supernate was removed, assayed for human ST2, and measured 29,920 pg/mL.

HUT-78 human cutaneous T cell lymphoma cells (1 x 104 cells/mL) were seeded in RPMI supplemented with 10% fetal bovine serum, 5 μM beta -mercaptoethanol, and 10 ng/mL recombinant human IL-2 until confluent. An aliquot of the cell culture supernate was removed, assayed for human ST2, and measured 54.9 pg/mL.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.

Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.

Cell Culture Supernates, EDTA Plasma, Heparin Plasma, Serum

	Intra-Assay Precision			Inter-Assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	40	40	40
Mean (pg/mL)	273	628	1027	262	642	1064
Standard Deviation	15.2	27.5	46.3	18.7	34.7	67.2
CV%	5.6	4.4	4.5	7.1	5.4	6.3

Recovery for Human ST2/IL-33R Quantikine ELISA Kit

The recovery of ST2 spiked to levels throughout the range of the assay was evaluated.

Sample Type	Average % Recovery	Range %
Cell Culture Media (n=4)	97	89-106

Linearity

To assess the linearity of the assay, samples containing or spiked with high concentrations of ST2 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.

Scientific Data Images for Human ST2/IL-33R Quantikine ELISA Kit

Human ST2/IL-1 R4 ELISA Standard Curve

Preparation and Storage

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: ST2/IL-33R

ST2, also known as IL-1R4, is a component of the receptor for the proinflammatory cytokine Interleukin-33. The IL-33 receptor additionally includes IL-1 RAcp which also associates with IL-1 RI and IL-1 R rp2/IL-1 R6. A soluble form of ST2 functions as a decoy receptor. IL-33 promotes Th2-biased immune responses, resulting in eosinophilia, mast cell activation, and allergic inflammation. In addition, IL-33 enhances inflammation-associated hypernociception and protects from atherosclerosis and cardiac myocyte hypertrophy. Soluble ST2 is elevated in the serum under inflammatory conditions including allergic asthma, sepsis, trauma, dengue fever, pulmonary disease, and lupus. It is also associated aortic stenosis, congestive cardiomyopathy, and risk of cardiovascular heart failure and death.

Long Name

Interleukin 33 Receptor

Alternate Names

DER4, Fit-1, IL-1 R4, IL-1R4, IL-1RL1, IL-33 R, IL-33R, IL1R4, IL33R, Ly84, ST2L, ST2V, T1

Entrez Gene IDs

9173 (Human); 17082 (Mouse)

Gene Symbol

IL1RL1

Additional ST2/IL-33R Products

Product Documents for Human ST2/IL-33R Quantikine ELISA Kit

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Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human ST2/IL-33R Quantikine ELISA Kit

For research use only

⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Related Research Areas

Group 2 ILCs

IL-1 Family

Citations for Human ST2/IL-33R Quantikine ELISA Kit

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Human ST2/IL-33 R Quantikine ELISA Kit

Name: Ivana Marventano

Sample Tested: Human serum

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Protocols

View specific protocols for Human ST2/IL-33R Quantikine ELISA Kit (DST200):

Refer to the product datasheet for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.

Prepare all reagents, standard dilutions, and samples as directed in the product insert.

Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

Add 50 µL of Assay Diluent to each well.

Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.

Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.

Aspirate and wash 4 times.

Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for Human ST2/IL-33R Quantikine ELISA Kit

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Q: Assay Diluent RD1-63 contains visible precipitate. Is this acceptable to use?

A: A small amount of precipitate is expected and acceptable in Assay Diluent RD1-63. Variability has been observed between vials and diluent lots. It may not fully resolubilize but this will not impact assay performance. It is not recommended to filter the precipitate.