Human LIF Quantikine ELISA Kit

R&D Systems | Catalog # DLF00

R&D Systems
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Key Product Details

Assay Length

3.5 hours or 4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (200 µL), Serum (200 µL), EDTA Plasma (200 µL), Heparin Plasma (200 µL)

Sensitivity

8 pg/mL

Assay Range

31.2-2000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
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Product Summary for Human LIF Quantikine ELISA Kit

The Quantikine Human LIF Immunoassay is a 3.5 or 4.5 hour solid phase ELISA designed to measure soluble human LIF in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human LIF and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant LIF accurately. Results obtained measuring natural human LIF showed dose-response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural human LIF.

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 LIF

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.

精度

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 20 20 20
Mean (pg/mL) 341 875 1358 81.6 472 910
Standard Deviation 12.3 34.5 42.5 4.4 17.7 35.2
CV% 3.6 3.9 3.1 5.4 3.8 3.9

EDTA Plasma, Heparin Plasma, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 314 868 1347 87.4 524 1017
Standard Deviation 7.7 24.3 23.8 6.2 34.9 46.5
CV% 2.5 2.8 1.8 7.1 6.7 4.6

Recovery for Human LIF Quantikine ELISA Kit

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

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 97 91-106
EDTA Plasma (n=2) 89 84-97
Heparin Plasma (n=8) 92 84-100
Serum (n=11) 102 91-110

Linearity

Human LIF ELISA Linearity

Scientific Data Images for Human LIF Quantikine ELISA Kit

Human LIF ELISA Cell Culture Supernate Standard Curve

Human LIF ELISA Cell Culture Supernate Standard Curve

Human LIF ELISA Standard Curve

Human LIF ELISA Standard Curve

Human LIF ELISA Serum/Plasma Standard Curve

Human LIF ELISA Serum/Plasma 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: LIF

LIF (Leukemia Inhibitory Factor) is a pleiotropic member of the IL-6 family of cytokines. It functions through a heterodimeric receptor complex containing a ligand-binding subunit, LIF R alpha/CD118, and a signal transducing subunit, gp130. gp130 also serves as a subunit of the receptor complexes for Oncostatin M, Cardiotrophin-1, CNTF, IL-6, IL-11, and IL-27. LIF supports uterine implantation of the embryo, pluripotency in embryonic stem cells, and proliferation of progenitor cells. It promotes regulatory T cell and inhibits Th17 cell differentiation, and it is required by the thymic epithelium to support T cell maturation. It promotes motor neuron survival and oligodendrocyte myelination, enhances adrenal production of cortisol and aldosterone, and promotes differentiation of adipocytes and cardiac smooth muscle cells. LIF can also function as an autocrine growth factor in pancreatic cancer and can induce formation of immunosuppressive tumor-associated macrophages.

Long Name

Leukemia Inhibitory Factor

Alternate Names

D Factor, Emfilermin, HILDA, MLPLI

Entrez Gene IDs

3976 (Human); 16878 (Mouse); 403449 (Canine)

Gene Symbol

LIF

Additional LIF Products

Product Documents for Human LIF 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 LIF 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 LIF Quantikine ELISA Kit

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Protocols

View specific protocols for Human LIF Quantikine ELISA Kit (DLF00):

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 for Serum & Plasma Samples Only
  4.   For Serum & Plasma Samples Only: Add 50 µL of Assay Diluent to each well.

  5. 200 µL Standard, Control, or Sample
  6.   Add 200 µL of Standard, control, or sample to each well.
  7.   For Serum & Plasma Samples: Cover with a plate sealer, and incubate at 37 °C for 2 hours.
    For Cell Culture Supernate Samples: Cover with a plate sealer, and incubate at room temperature for 2 hours.
  8.   Aspirate each well and wash, repeating the process twice for a total of 3 washes.

  9. 200 µL Conjugate
  10.   Add 200 µL of Conjugate to each well.
  11.   For Serum & Plasma Samples: Cover with a new plate sealer, and incubate at room temperature for 2 hours.
    For Cell Culture Supernate Samples: Cover with a new plate sealer, and incubate at room temperature for 1 hour.
  12.   Aspirate and wash 3 times.

  13. 200 µL Substrate Solution
  14. Add 200 µL Substrate Solution to each well. Incubate at room temperature for 20 minutes. PROTECT FROM LIGHT.

  15. 50 µL Stop Solution
  16. 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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