Human S100A8/S100A9 Heterodimer DuoSet ELISA

R&D Systems | Catalog # DY8226-05

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Key Product Details

Assay Type

Solid Phase Sandwich ELISA

Assay Range

93.8-6000 pg/mL

Sample Type

Cell culture supernates, serum, and plasma
Note: Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet

Reactivity

Human

Human S100A8/S100A9 Heterodimer DuoSet ELISA Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits
View all S100A8/S100A9 Heterodimer ELISA Kits »

Product Summary for Human S100A8/S100A9 Heterodimer DuoSet ELISA

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and Recombinant human S100 Calcium Binding Protein A8/S100 Calcium Binding Protein A9 (S100A8/S100A9 Heterodimer). The suggested diluent is suitable for the analysis of most cell culture supernate, serum, plasma, saliva, human milk, urine, and fecal samples. Diluents for complex matrices, such as serum, plasma, saliva, human milk, urine, and feces, should be evaluated prior to use in this DuoSet.

Product Specifications

Assay Format

96-well strip plate (sold separately)

Detection Method

Colorimetric ELISA - 450nm (TMB)

Conjugate

Biotin

Specificity

Please see the product datasheet

Label

HRP

Scientific Data Images for Human S100A8/S100A9 Heterodimer DuoSet ELISA

Human S100A8 / S100A9 Heterodimer ELISA Standard Curve

Human S100A8 / S100A9 Heterodimer ELISA Standard Curve

Kit Contents for Human S100A8/S100A9 Heterodimer DuoSet ELISA

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

PBS:

(Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or equivalent

Reagent Diluent*

Blocking Buffer*

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990), or equivalent

Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent

*For the Reagent Diluent and Blocking Buffer recommended for a specific DuoSet ELISA Development Kit, please see the product.

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: S100A8/S100A9 Heterodimer

S100A8 (also known as MRP8, Calgranulin A, and CP-10) and S100A9 (also known as MRP14 and Calgranulin B) are secreted pro-inflammatory proteins that are upregulated in neutrophils and monocytes at sites of inflammation (e.g. psoriasis, rheumatoid arthritis, cardiac ischemia) and are present at elevated concentrations in rheumatoid arthritis synovial fluid. In the presence of calcium or zinc, S100A8 and S100A9 associate into noncovalent homodimers and heterodimers with each other. The heterodimer additionally binds and sequesters manganese, thereby restricting the growth of Mn-dependent bacteria. The S100A8/A9 heterodimer exhibits functions beyond those performed by the individual proteins. These include binding to fatty acids such as arachidonic acid and promoting astrocyte proliferation. S100A8, S100A9, and the heterodimer each promote neutrophil infiltration into sites of inflammation and inflammatory cytokine production by monocytes.

Long Name

S100 Calcium Binding Protein A8/S100 Calcium Binding Protein A9

Alternate Names

Calprotectin

Entrez Gene IDs

6279 (Human)

Gene Symbol

S100A8

Additional S100A8/S100A9 Heterodimer Products

Product Documents for Human S100A8/S100A9 Heterodimer DuoSet ELISA

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 S100A8/S100A9 Heterodimer DuoSet ELISA

For research use only

Citations for Human S100A8/S100A9 Heterodimer DuoSet ELISA

Customer Reviews for Human S100A8/S100A9 Heterodimer DuoSet ELISA (4)

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  • Human S100A8/S100A9 Heterodimer DuoSet ELISA
    Name: Anonymous
    Sample Tested: Cell culture supernatant
    Verified Customer | Posted 11/19/2021
    Human S100A8/S100A9 Heterodimer DuoSet ELISA DY8226-05
  • Human S100A8/S100A9 Heterodimer DuoSet ELISA
    Name: Anonymous
    Sample Tested: Cell culture supernatant
    Verified Customer | Posted 09/14/2021
  • Human S100A8/S100A9 Heterodimer DuoSet ELISA
    Name: Anonymous
    Sample Tested: Human keratinocytes
    Verified Customer | Posted 12/16/2019
    Human S100A8/S100A9 Heterodimer DuoSet ELISA DY8226-05
  • Human S100A8/S100A9 Heterodimer DuoSet ELISA
    Name: Anonymous
    Sample Tested: Serum
    Verified Customer | Posted 10/11/2018

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Protocols

View specific protocols for Human S100A8/S100A9 Heterodimer DuoSet ELISA (DY8226-05):

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Block Buffer to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent with NGS, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

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FAQs for Human S100A8/S100A9 Heterodimer DuoSet ELISA

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    • Q: For the Human S100A8/S100A9 Heterodimer DuoSet ELISA, Catalog # DY8226-05, can Tween 20 be substituted for Brij® 35 in the Reagent Diluent?
      A: The 50 mM Tris, 10 mM CaCl2, 0.15 M NaCl, 0.05% Brij® 35, pH 7.45-7.5 that we recommend as a reagent diluent for this kit is unique, which means that our lab would have found that more traditional reagent diluents do not perform optimally. It is certainly possible that you could potentially substitute a different detergent (such as Tween) with viable results, but the performance of this different formulation of the reagent diluent would need to be empirically determined on your end.
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