Rat IL-6 DuoSet ELISA

R&D Systems | Catalog # DY506

R&D Systems
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Key Product Details

Assay Type

Solid Phase Sandwich ELISA

Assay Range

125-8000 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

Rat

Rat IL-6 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
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Product Summary for Rat IL-6 DuoSet ELISA

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant rat IL-6. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Specifications

Assay Format

96-well strip plate (sold separately)

Sample Volume Required

100 µL

Detection Method

Colorimetric ELISA - 450nm (TMB)

Conjugate

Biotin

Label

HRP

Scientific Data Images for Rat IL-6 DuoSet ELISA

Rat IL-6 ELISA Standard Curve

Rat IL-6 ELISA Standard Curve

Kit Contents for Rat IL-6 DuoSet ELISA

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

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

Normal Goat Serum: (Catalog # DY005)

The components listed above may be purchased separately:

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 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

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)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Normal Goat Serum: (Catalog # DY005)

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-6

IL-6 (Interleukin-6) is a pleiotropic cytokine that acts in the acute phase reaction, inflammation, hematopoiesis, bone metabolism, and cancer progression. It contributes to chronic inflammation in obesity, inflammatory bowel disease, arthritis, sepsis, and atherosclerosis. IL-6 signals through a receptor complex of IL-6 R alpha and gp130. gp130 is also a component of the receptors for CLC, CNTF, CT-1, IL-11, IL-27, LIF, and OSM. Soluble forms of IL-6 R alpha are generated by both alternative splicing and proteolytic cleavage. In a mechanism known as trans-signaling, complexes of soluble IL-6 and IL-6 R alpha elicit responses from gp130-expressing cells that lack cell surface IL-6 R alpha.


R&D Systems, a Bio-Techne brand, offers a comprehensive range of IL-6 Antibody products, IL-6 Proteins and Enzymes, IL-6 ELISAs, IL-6 Luminex Assays and more. Use the filters above to drill down to the IL-6 product that best fits your needs.

Long Name

Interleukin 6

Alternate Names

BSF-2, BSF2, IFNB2, IL6, MGI-2A

Entrez Gene IDs

3569 (Human); 16193 (Mouse); 24498 (Rat); 399500 (Porcine); 280826 (Bovine); 403985 (Canine); 102138971 (Cynomolgus Monkey); 100034196 (Equine); 493687 (Feline); 463288 (Primate); 100008733 (Rabbit)

Gene Symbol

IL6

Additional IL-6 Products

Product Documents for Rat IL-6 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 Rat IL-6 DuoSet ELISA

For research use only

Citations for Rat IL-6 DuoSet ELISA

Customer Reviews for Rat IL-6 DuoSet ELISA (7)

4.3 out of 5
7 Customer Ratings
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  • Rat IL-6 DuoSet ELISA
    Name: Anonymous
    Sample Tested: rat lung
    Verified Customer | Posted 10/05/2023
    Rat IL-6 DuoSet ELISA DY506
  • Rat IL-6 DuoSet ELISA
    Name: Anonymous
    Sample Tested: Lung tissue
    Verified Customer | Posted 08/01/2023
    Rat IL-6 DuoSet ELISA DY506
  • Rat IL-6 DuoSet ELISA
    Name: Anonymous
    Sample Tested: Colon tissue
    Verified Customer | Posted 05/19/2023
    Rat IL-6 DuoSet ELISA DY506
  • Rat IL-6 DuoSet ELISA
    Name: Anonymous
    Sample Tested: Peritoneal fluid
    Verified Customer | Posted 02/11/2023
    Rat IL-6 DuoSet ELISA DY506
  • Rat IL-6 DuoSet ELISA
    Name: Anonymous
    Sample Tested: Serum, broncho-alveloar lavage fluid and Tissue Homogenates
    Verified Customer | Posted 08/11/2020
    Rat IL-6 DuoSet ELISA DY506
  • Name: Sebastian Halbach
    Sample Tested: RBL-1 rat basophilic leukemia cell line
    Verified Customer | Posted 10/31/2019
  • Rat IL-6 DuoSet ELISA
    Name: Anonymous
    Sample Tested: Cell Culture Supernates and Plasma
    Verified Customer | Posted 08/21/2016

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Protocols

View specific protocols for Rat IL-6 DuoSet ELISA (DY506):

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 Reagent Diluent 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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