Human LIF Quantikine ELISA Kit

  (17 citations)     
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Product Details
Assay Procedure
Citations (17)
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  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    3.5 hours or 4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (200 uL), Serum (200 uL), EDTA Plasma (200 uL), Heparin Plasma (200 uL)
  • Sensitivity
    8 pg/mL
  • Assay Range
    31.2 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
  • 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.
Control Available
QC113 , Quantikine Immunoassay Control Set 582 for Human LIF - Please Inquire
Product Summary
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.

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 20 20 20
Mean 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

Serum, EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 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

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
Product Datasheets

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Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: LIF
Based on its helical structure, LIF (Leukemia Inhibitory Factor) is considered a member of the Interleukin-6 family of cytokines. Functionally, it has been implicated in a many physiological processes including development, hematopoiesis, bone metabolism, and inflammation. Some cell types known to express LIF include activated T cells, monocytes, astrocytes, osteoblasts, keratinocytes, regenerating skeletal muscle, mast cells, and fibroblasts. The activities of LIF are mediated through a high-affinity heterodimeric receptor complex consisting of two membrane glycoproteins: an alpha subunit (LIF R alpha, also known as LIF R beta and CD118) that binds LIF with low affinity and the 130 kDa (gp130) subunit that does not bind LIF by itself, but is required for high-affinity binding of LIF by the complex.
  • Long Name:
    Leukemia Inhibitory Factor
  • Entrez Gene IDs:
    3976 (Human); 16878 (Mouse); 403449 (Canine)
  • Alternate Names:
    CDF; D factor; DIA; differentiation inhibitory activity; differentiation stimulating factor; Differentiation-stimulating factor; Emfilermin; HILDAcholinergic differentiation factor; leukemia inhibitory factor (cholinergic differentiation factor); leukemia inhibitory factor; LIF; Melanoma-derived LPL inhibitor; MLPLI
Related Research Areas
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 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.
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

17 Citations: Showing 1 - 10
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Species
Sample Type
  1. The role of p38 mitogen-activated protein kinase in serum-induced leukemia inhibitory factor secretion by bone marrow stromal cells from pediatric myelodysplastic syndromes.
    Authors: da Costa SV, Roela RA, Junqueira MS, Arantes C, Brentani MM
    Leuk. Res., 2010;34(4):507-12.
    Species: Human
    Sample Type: Cell Culture Supernates
  2. Leukemia inhibitory factor is dysregulated in the endometrium and uterine flushing fluid of patients with adenomyosis during implantation window.
    Authors: Xiao Y, Sun X, Yang X, Zhang J, Xue Q, Cai B, Zhou Y
    Fertil. Steril., 2010;94(1):85-9.
    Species: Human
    Sample Type: Uterine Lavage Fluid
  3. Prokineticin 1 mediates fetal-maternal dialogue regulating endometrial leukemia inhibitory factor.
    Authors: Evans J, Catalano RD, Brown P, Sherwin R, Critchley HO, Fazleabas AT, Jabbour HN
    FASEB J., 2009;23(7):2165-75.
    Species: Human
    Sample Type: Cell Culture Supernates
  4. Helicobacter pylori cytotoxin-associated gene A activates the signal transducer and activator of transcription 3 pathway in vitro and in vivo.
    Authors: Bronte-Tinkew DM, Terebiznik M, Franco A, Ang M, Ahn D, Mimuro H, Sasakawa C, Ropeleski MJ, Peek RM, Jones NL
    Cancer Res., 2009;69(2):632-9.
    Species: Human
    Sample Type: Cell Culture Supernates
  5. Isoflavones regulate secretion of leukemia inhibitory factor and transforming growth factor {beta} and expression of glycodelin in human endometrial epithelial cells.
    Authors: Xu JW, Yasui N, Ikeda K, Pan WJ, Watanabe J, Shiotani M, Yanaihara A, Miki T, Yamori Y
    J. Endocrinol., 2008;196(2):425-33.
    Species: Human
    Sample Type: Cell Culture Supernates
  6. Tumor-associated leukemia inhibitory factor and IL-6 skew monocyte differentiation into tumor-associated macrophage-like cells.
    Authors: Duluc D, Delneste Y, Tan F, Moles MP, Grimaud L, Lenoir J, Preisser L, Anegon I, Catala L, Ifrah N, Descamps P, Gamelin E, Gascan H, Hebbar M, Jeannin P
    Blood, 2007;110(13):4319-30.
    Species: Human
    Sample Type: Cell Culture Supernates
  7. Leukemia inhibitory factor functions as a growth factor in pancreas carcinoma cells: Involvement of regulation of LIF and its receptor expression.
    Authors: Kamohara H, Ogawa M, Ishiko T, Sakamoto K, Baba H
    Int. J. Oncol., 2007;30(4):977-83.
    Species: Human
    Sample Type: Cell Culture Supernates
  8. Increased ENA-78 in the follicular fluid of patients with endometriosis.
    Authors: Wunder DM, Mueller MD, Birkhauser MH, Bersinger NA
    Acta Obstet Gynecol Scand, 2006;85(3):336-42.
    Species: Human
    Sample Type: Follicular Fluid
  9. Interleukin-11 potentiates oligodendrocyte survival and maturation, and myelin formation.
    Authors: Zhang Y, Taveggia C, Melendez-Vasquez C, Einheber S, Raine CS, Salzer JL, Brosnan CF, John GR
    J. Neurosci., 2006;26(47):12174-85.
    Species: Human
    Sample Type: Cell Culture Supernates
  10. Circulating interleukin-6 family cytokines and their receptors in patients with congestive heart failure.
    Authors: Hirota H, Izumi M, Hamaguchi T, Sugiyama S, Murakami E, Kunisada K, Fujio Y, Oshima Y, Nakaoka Y, Yamauchi-Takihara K
    Heart Vessels, 2004;19(5):237-41.
    Species: Human
    Sample Type: Plasma
  11. Effect of azacytidine in the release of leukemia inhibitory factor, oncostatin m, interleukin (IL)-6, and IL-11 by mononuclear cells of patients with refractory anemia.
    Authors: Lopez-Karpovitch X, Barrales-Benitez O, Flores M, Piedras J
    Cytokine, 2002;20(4):154-62.
    Species: Human
    Sample Type: Cell Culture Supernates
  12. Role of mitogen-activated protein kinase family in serum-induced leukaemia inhibitory factor and interleukin-6 secretion by bone marrow stromal cells.
    Authors: Nakao T, Kim S, Ohta K, Kawano H, Hino M, Miura K, Tatsumi N, Iwao H
    Br. J. Pharmacol., 2002;136(7):975-84.
    Species: Human
    Sample Type: Cell Culture Supernates
  13. The significance of cytokines, chemical composition, and murine embryo development in hydrosalpinx fluid for predicting the IVF outcome in women with hydrosalpinx.
    Authors: Chen CD, Yang JH, Lin KC, Chao KH, Ho HN, Yang YS
    Hum. Reprod., 2002;17(1):128-33.
    Species: Human
    Sample Type: Hydrosalpinx Fluid
  14. Distinct hematopoietic support by two human stromal cell lines.
    Authors: Loeuillet C, Bernard G, Remy-Martin J, Saas P, Herve P, Douay L, Chalmers D
    Exp. Hematol., 2001;29(6):736-45.
    Species: Human
    Sample Type: Cell Culture Supernates
  15. Expression of interleukin-6, leukemia inhibitory factor and their receptors by colonic epithelium and pericryptal fibroblasts.
    Authors: Rockman SP, Demmler K, Roczo N, Cosgriff A, Phillips WA, Thomas RJ, Whitehead RH
    J. Gastroenterol. Hepatol., 2001;16(9):991-1000.
    Species: Human
    Sample Type: Cell Culture Supernates
  16. Interleukin-6 is a major effector molecule of short-term G-CSF treatment inducing bone metabolism and an acute-phase response.
    Authors: Carstanjen D, Regenfus M, Muller C, Salama A
    Exp. Hematol., 2001;29(7):812-21.
    Species: Human
    Sample Type: Plasma
  17. Influence of protein supplements on the secretion of leukaemia inhibitory factor by mitomycin-pretreated Vero cells: possible application to the in vitro production of bovine blastocysts with high cryotolerance.
    Authors: Carnegie JA, Morgan JJ, McDiarmid N, Durnford R
    J. Reprod. Fertil., 1999;117(1):41-8.
    Species: Primate - Cholrocebus pygerythrus (Vervet Monkey)
    Sample Type: Cell Culture Supernates
Expand to show all Citations

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Quantikine Immunoassay Control Set 582 for Human LIF

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