Mouse IL-6 R alpha Quantikine ELISA Kit

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MR600
Mouse IL-6 R alpha Standard Curve
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Mouse IL-6 R alpha Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (10 uL), Heparin Plasma (10 uL)
Sensitivity
0.07 ng/mL
Assay Range
0.2 - 12 ng/mL (Cell Culture Supernates, Serum, Heparin Plasma)
Specificity
Natural and recombinant mouse IL-6 R alpha.
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.

Product Summary

The Quantikine Mouse IL-6 R alpha Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse IL-6 R alpha levels in cell culture supernates, serum, and heparin plasma. It contains NS0-expressed recombinant mouse IL-6 R alpha and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant mouse IL-6 R alpha accurately. Results obtained using natural mouse IL-6 R alpha showed dose-response curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse IL-6 R alpha.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components

Cell Culture Supernates, Serum, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 0.456 1.07 2.94 0.474 1.08 2.82
Standard Deviation 0.036 0.039 0.148 0.06 0.08 0.2
CV% 7.9 3.6 5 12.7 7.4 7.1

Recovery

The recovery of mouse IL-6 R 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) 98 90-110
Heparin Plasma (n=4) 99 92-106
Serum (n=4) 102 93-110

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of mouse IL-6 R alpha were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Mouse IL-6 R alpha ELISA Linearity

Data Examples

Mouse IL-6 R alpha Standard Curve

Product Datasheets

Preparation and Storage

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

Background: IL-6 R alpha

IL-6 R alpha is a transmembrane subunit of the receptor for Interleukin-6. It associates with gp130 which is a shared 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. IL-6 plays important roles in the acute phase reaction, inflammation, hematopoiesis, bone metabolism, and cancer progression. IL-6, along with TNF-alpha and IL-1, drives the acute inflammatory response and the transition from acute inflammation to either acquired immunity or chronic inflammatory disease. When dysregulated, it contributes to chronic inflammation in obesity, insulin resistance, inflammatory bowel disease, arthritis, sepsis, and atherosclerosis.

Long Name:
Interleukin 6 Receptor alpha
Entrez Gene IDs:
3570 (Human); 16194 (Mouse)
Alternate Names:
CD126; gp80; IL-6 R alpha; IL6Q; IL6R alpha; IL6R; IL-6R-1; IL6RA; IL-6Ra; IL6RQ; interleukin 6 receptor
⚠ 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.

Assay Procedure

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
  4.   Add 50 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 100 µL Conjugate
  9.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
  10.   Aspirate and wash 4 times.

  11. 100 µL Substrate Solution
  12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  13. 100 µL Stop Solution
  14.   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.

FAQs

No product specific FAQs exist for this product, however you may

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Mouse IL-6 R alpha Quantikine ELISA Kit
By Anonymous on 09/30/2016
Sample Tested: Serum and Plasma

Great kit.