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Human IL-1 alpha/IL-1F1 Quantikine ELISA Kit

R&D Systems | Catalog # DLA50

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Key Product Details

Assay Length

3.5 hours or 4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (200 µL), Serum (200 µL), EDTA Plasma (200 µL), Heparin Plasma (200 µL), Citrate Plasma (200 µL)

Sensitivity

1 pg/mL

Assay Range

3.9-250 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
View all IL-1 alpha/IL-1F1 ELISA Kits »

Product Summary for Human IL-1 alpha/IL-1F1 Quantikine ELISA Kit

The Quantikine Human IL-1 alpha Immunoassay is a 3.5 or 4.5 hour solid phase ELISA designed to measure IL-1 alpha in cell culture supernates, serum, and plasma. It contains recombinant human IL-1 alpha and antibodies raised against recombinant human IL-1 alpha and has been shown to accurately quantitate the recombinant factor. Results obtained using natural IL-1 alpha showed linear curves that were parallel to the standard curves obtained using the E. coli-expressed Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural human IL-1 alpha.

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 IL-1 alpha

Cross-reactivity

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

Interference

Interference observed with 1 or more available related molecules.

Sample Values

Serum/Plasma - Forty samples from apparently healthy volunteers were evaluated for the presence of human IL-1 alpha, and all had levels which fell below the lowest IL-1 alpha standard, 3.9 pg/mL. No medical histories were available for the donors used in this study.

Cell Culture Supernates - Human peripheral blood mononuclear cells (5 x 106 cells/mL) were cultured in RPMI supplemented with 5% fetal bovine serum, 50 μM beta -mercaptoethanol, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. Cells were cultured unstimulated or stimulated with 10 μg/mL PHA. Aliquots of the cell culture supernates were removed on days 1 and 5 and assayed for levels of human IL-1 alpha.

ConditionDay 1 (pg/mL)Day 5 (pg/mL)
Unstimulated8523
Stimulated1330395

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

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 26.9 122 191 20.5 60.0 121
Standard Deviation 0.4 4.6 6.6 1.7 3.1 5.2
CV% 1.5 3.8 3.5 8.3 5.2 4.3

Citrate Plasma, EDTA Plasma, Heparin Plasma, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 28.6 125 185 27.9 123 187
Standard Deviation 0.4 1.4 4.0 1.2 4.7 6.3
CV% 1.4 1.1 2.2 4.3 3.8 3.4

Recovery for Human IL-1 alpha/IL-1F1 Quantikine ELISA Kit

The recovery of IL-1 alpha spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 96 91-104
Citrate Plasma (n=6) 97 71-119
EDTA Plasma (n=6) 104 84-121
Heparin Plasma (n=6) 104 72-126
Serum (n=6) 107 74-127

Linearity

To assess the linearity of the assay, the following biological samples spiked with high concentrations of IL-1 alpha were diluted with the appropriate Calibrator Diluent and then assayed.

Human IL-1 alpha/IL-1F1 ELISA Linearity

Scientific Data Images for Human IL-1 alpha/IL-1F1 Quantikine ELISA Kit

Human IL-1 alpha/IL-1F1 ELISA Calibrator Diluent RD5-5 Standard Curve

Human IL-1 alpha/IL-1F1 ELISA Calibrator Diluent RD5-5 Standard Curve

Human IL-1 alpha/IL-1F1 ELISA Calibrator Diluent RD6C Standard Curve

Human IL-1 alpha/IL-1F1 ELISA Calibrator Diluent RD6C 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: IL-1 alpha/IL-1F1

IL-1 alpha (Interleukin-1 alpha) is a cytokine that is induced during microbial exposure and promotes the release of multiple inflammatory mediators and acute phase proteins. It also supports hematopoietic stem cell expansion, promotes neutrophil mobilization from the bone marrow, and promotes osteoclast formation. IL-1 alpha signals through a receptor complex composed of IL-1 RI and IL-1 RAcP. It also binds the decoy receptor IL-1 RII which associates with IL-1 RAcP but does not transduce signalling.

 

Long Name

Interleukin 1 alpha

Alternate Names

BAF, IL-1A, IL-1F1, IL1, IL1 alpha, IL1A, IL1F1, LAF, LEM

Entrez Gene IDs

3552 (Human); 16175 (Mouse); 24493 (Rat); 397094 (Porcine); 101926546 (Cynomolgus Monkey)

Gene Symbol

IL1A

Additional IL-1 alpha/IL-1F1 Products

Product Documents for Human IL-1 alpha/IL-1F1 Quantikine ELISA Kit

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.

Product Specific Notices for Human IL-1 alpha/IL-1F1 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.

Citations for Human IL-1 alpha/IL-1F1 Quantikine ELISA Kit

Customer Reviews for Human IL-1 alpha/IL-1F1 Quantikine ELISA Kit (9)

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  • Human IL-1 alpha/IL-1F1 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Serum-free Cell Culture Media
    Verified Customer | Posted 06/08/2022
  • Human IL-1 alpha/IL-1F1 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Human cell conditioned medium
    Verified Customer | Posted 08/31/2021
  • Human IL-1 alpha/IL-1F1 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: conditioned media
    Verified Customer | Posted 02/28/2021
    Human IL-1 alpha/IL-1F1 Quantikine ELISA Kit DLA50
  • Human IL-1 alpha/IL-1F1 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Serum
    Verified Customer | Posted 08/06/2018
  • Human IL-1 alpha/IL-1F1 Quantikine ELISA Kit
    Name: Alisha Freeman
    Sample Tested: A-673 human Ewing's sarcoma cell line
    Verified Customer | Posted 07/11/2018
  • Human IL-1 alpha/IL-1F1 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Pancreatic cancer cells
    Verified Customer | Posted 06/27/2018
  • Human IL-1 alpha/IL-1F1 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Adipocytes
    Verified Customer | Posted 04/11/2018
  • Human IL-1 alpha/IL-1F1 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: PANC-1 human pancreatic carcinoma cell line
    Verified Customer | Posted 03/30/2018
    Human IL-1 alpha/IL-1F1 Quantikine ELISA Kit DLA50
  • Human IL-1 alpha/IL-1F1 Quantikine ELISA Kit
    Name: Alisha Freeman
    Sample Tested: CHO Chinese hamster ovary cell line
    Verified Customer | Posted 12/07/2017

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Protocols

View specific protocols for Human IL-1 alpha/IL-1F1 Quantikine ELISA Kit (DLA50):

Refer to the product 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.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
    3. 50 µL Assay Diluent
  3.   Add 50 µL of Assay Diluent to each well.
    4. 200 µL Standard, Control, or Sample
  4.   Add 200 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  5.   Aspirate each well and wash, repeating the process twice for a total of 3 washes.
    6. 200 µL Conjugate
  6.   Add 200 µL of Conjugate to each well.
  7.   For Cell Culture Supernate Samples: Cover with a new plate sealer, and incubate at room temperature for 1 hour.
    For Serum & Plasma Samples: Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  8.   Aspirate and wash 3 times.
    9. 200 µL Substrate Solution
  9.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 20 minutes. PROTECT FROM LIGHT.
    10. 50 µL Stop Solution
  10. Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

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