Human Soluble gp130 Quantikine ELISA Kit

R&D Systems | Catalog # DGP00

R&D Systems
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Key Product Details

Assay Length

4.5 hours or 5.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (100 µL), Serum (10 µL), EDTA Plasma (10 µL), Heparin Plasma (10 µL), Citrate Plasma (10 µL)

Sensitivity

0.05 ng/mL (Cell Culture Supernates)
0.08 ng/mL (Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)

Assay Range

0.125-8 ng/mL (Cell Culture Supernates)
0.25-16 ng/mL (Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
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Product Summary for Human Soluble gp130 Quantikine ELISA Kit

The Quantikine Human soluble gp130 Immunoassay is a 4.5 or 5.5 hour solid phase ELISA designed to measure sgp130 in cell culture supernates, serum, and plasma. It contains soluble insect cell-expressed recombinant human gp130 and antibodies raised against recombinant human gp130 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural gp130 showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural gp130.

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 sgp130

Cross-reactivity

< 0.5% cross-reactivity observed with available related molecules. Cross-species reactivity not 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) 0.40 1.00 4.43 0.40 1.02 4.50
Standard Deviation 0.02 0.07 0.33 0.02 0.10 0.31
CV% 5.0 7.0 7.4 5.0 9.8 6.9

Citrate Plasma, EDTA Plasma, Heparin Plasma, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 0.70 1.82 8.43 0.77 1.94 8.64
Standard Deviation 0.03 0.10 0.40 0.04 0.07 0.30
CV% 4.3 5.5 4.7 5.2 3.6 3.5

Recovery for Human Soluble gp130 Quantikine ELISA Kit

The recovery of sgp130 spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=5) 109 96-114
Citrate Plasma (n=5) 98 84-112
EDTA Plasma (n=5) 100 83-114
Heparin Plasma (n=5) 98 85-110
Serum (n=5) 101 83-120

Linearity

To assess the linearity of the assay, five samples were spiked with high concentrations of sgp130 in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.

Human gp130 ELISA Linearity

Scientific Data Images for Human Soluble gp130 Quantikine ELISA Kit

Human gp130 ELISA Serum/Plasma Standard Curve

Human gp130 ELISA Serum/Plasma Standard Curve

Human gp130 ELISA Cell Culture Supernate Standard Curve

Human gp130 ELISA Cell Culture Supernate 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: gp130

Gp130, the common signal transducing receptor component shared by the functional receptor complexes of the IL-6 family of cytokines, belongs to the classI cytokine receptor family. Binding of IL-6 or IL-11 to either the membrane-anchored or soluble IL-6 R or IL-11 R, respectively, initiates the association of IL-6 R (IL-11 R) with gp130. The complex then undergoes homodimerization and signal transduction. For other IL-6 family cytokines, such as LIF and OSM, signal transduction is similarly triggered by the heterodimerization of gp130 with LIFR or OSM R.

Long Name

Glycoprotein 130

Alternate Names

CD130, IL6ST

Entrez Gene IDs

3572 (Human); 16195 (Mouse); 25205 (Rat); 101926138 (Cynomolgus Monkey)

Gene Symbol

IL6ST

Additional gp130 Products

Product Documents for Human Soluble gp130 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 Soluble gp130 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 Soluble gp130 Quantikine ELISA Kit

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  • Human Soluble gp130 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: A375s2 human melanoma cell line, subclone 2
    Verified Customer | Posted 01/07/2022
    Human Soluble gp130 Quantikine ELISA Kit DGP00
  • Human Soluble gp130 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Serum and Plasma and Cell culture supernatant
    Verified Customer | Posted 01/07/2020
  • Human Soluble gp130 Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Cell Culture Supernates
    Verified Customer | Posted 11/07/2019
    Human Soluble gp130 Quantikine ELISA Kit DGP00
  • Human Soluble gp130 Quantikine ELISA Kit
    Name: Jessica Fernando
    Sample Tested: Serum
    Verified Customer | Posted 12/04/2017

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Protocols

View specific protocols for Human Soluble gp130 Quantikine ELISA Kit (DGP00):

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. 100 µL Assay Diluent
  3.   Add 100 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 3 hours.
  5.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    6. 200 µL Conjugate
  6.   Add 200 µL of Conjugate to each well.
  7.   For Serum & Plasma Samples: Cover with a new plate sealer, and incubate at room temperature for 1 hour.
    For Cell Culture Supernate Samples: Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  8.   Aspirate and wash 4 times.
    9. 200 µL Substrate Solution
  9.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 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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