Human Caspase-1/ICE Quantikine ELISA Kit

R&D Systems | Catalog # DCA100

R&D Systems
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Key Product Details

Assay Length

3 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (100 µL)

Sensitivity

1.24 pg/mL

Assay Range

6.25-400 pg/mL (Cell Culture Supernates)
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Product Summary for Human Caspase-1/ICE Quantikine ELISA Kit

The Quantikine Human Caspase-1 Immunoassay is a 3 hour solid phase ELISA designed to measure Caspase-1 in cell culture supernates. It contains recombinant human Caspase-1 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained measuring natural human Caspase-1 showed dose-response curves that were parallel to the standard curves obtained using the recombinant Caspase-1. These results indicate that this kit can be used to determine relative mass values for natural human Caspase-1.

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 Caspase-1

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.

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 40 40 40
Mean (pg/mL) 41.0 77.7 228 40.4 75.2 219
Standard Deviation 2.49 4.62 11.11 3.8 6.9 18.1
CV% 6.1 5.9 4.9 9.4 9.2 8.3

Recovery for Human Caspase-1/ICE Quantikine ELISA Kit

The recovery of Caspase-1 spiked to three different levels throughout the range of the assay was evaluated

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 93 85-106

Linearity

To assess the linearity of the assay, cell culture supernate samples containing and/or spiked with high concentrations of Caspase-1 were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.

Human Caspase-1 ELISA Linearity

Scientific Data Images for Human Caspase-1/ICE Quantikine ELISA Kit

Human Caspase-1 ELISA Standard Curve

Human Caspase-1 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: Caspase-1

Caspase-1, also known as IL-1 beta-converting enzyme (ICE), is an aspartic protease that plays a key role in the inflammatory response and apoptosis. Caspase-1 precursor (about 50kDa) can be cleaved and the active enzyme consists of a complex of two 20 kDa and two 10 kDa subunits which associate following cleavage of inactive precursors. Caspase-1 is required for proteolytic cleavage of the IL-1 beta precursor to form the active proinflammatory cytokine.

 

Alternate Names

CASP1, Caspase1, ICE

Entrez Gene IDs

834 (Human)

Gene Symbol

CASP1

Additional Caspase-1 Products

Product Documents for Human Caspase-1/ICE 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 Caspase-1/ICE 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 Caspase-1/ICE Quantikine ELISA Kit

Customer Reviews for Human Caspase-1/ICE Quantikine ELISA Kit (3)

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  • Human Caspase-1/ICE Quantikine ELISA Kit
    Name: Phoibe Renema
    Sample Tested: Human Plasma
    Verified Customer | Posted 12/02/2017
  • Human Caspase-1/ICE Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Cell Culture Supernates
    Verified Customer | Posted 06/01/2017
    Human Caspase-1/ICE Quantikine ELISA Kit DCA100
  • Human Caspase-1/ICE Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Human plasma EDTA
    Verified Customer | Posted 05/10/2016
    Human Caspase-1/ICE Quantikine ELISA Kit DCA100

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Protocols

View specific protocols for Human Caspase-1/ICE Quantikine ELISA Kit (DCA100):

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. 100 µL Standard, Control, or Sample
  4.   Add 100 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 1.5 hours.
  5.   Aspirate each well and wash, repeating the process twice for a total of 3 washes.
    6. 100 µL Antiserum
  6.   Add 100 µL of Antiserum to each well. Cover with a new plate sealer, and incubate at room temperature for 30 minutes.
  7.   Aspirate and wash 3 times.
    8. 100 µL Conjugate
  8.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 30 minutes.
  9.   Aspirate and wash 3 times.
    10. 200 µL Substrate Solution
  10. Add 200 µL Substrate Solution to each well. Incubate at room temperature for 20 minutes. PROTECT FROM LIGHT.
    11. 50 µL Stop Solution
  11. 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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