Human SCF Quantikine ELISA Kit

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Human SCF/c-kit Ligand ELISA Cell Culture Supernate Standard Curve
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Human SCF 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 (100 uL), Serum (100 uL), EDTA Plasma (100 uL), Heparin Plasma (100 uL), Citrate Plasma (100 uL)
Sensitivity
9 pg/mL
Assay Range
31.2 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
Specificity
Natural and recombinant human SCF. This assay cross-reacts 15.6% with Canine SCF, 16.1% with Feline SCF, 3.5% with Mouse SCF and 1.0% with Rat SCF.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity observed with 1 or more species tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine Human SCF Immunoassay is a 4.5 hour solid phase ELISA designed to measure SCF levels in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human soluble SCF (KL-1 variant) and antibodies raised against this recombinant factor. Results obtained for naturally occurring human SCF samples 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 human SCF.

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 (pg/mL) 153 709 1427 151 729 1552
Standard Deviation 3.6 11.6 61 12.1 45.8 125
CV% 2.4 1.6 4.3 8 6.3 8.1

Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 139 655 1419 142 703 1574
Standard Deviation 5.5 12.9 54 14.6 36 94
CV% 4 2 3.8 10.3 5.1 6

Recovery

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

Sample Type Average % Recovery Range %
Cell Culture Media (n=2) 85 75-93
Citrate Plasma (n=8) 94 84-112
EDTA Plasma (n=8) 98 82-110
Heparin Plasma (n=8) 99 88-116
Serum (n=8) 99 87-111

Linearity

To assess the linearity of the assay, the following biological samples were diluted with the appropriate Calibrator Diluent and then assayed. The cell culture media was spiked with exogenous SCF.
Human SCF/c-kit Ligand ELISA Linearity

Scientific Data

Human SCF/c-kit Ligand ELISA Cell Culture Supernate Standard Curve

Human SCF/c-kit Ligand ELISA Serum/Plasma Standard Curve

Product Datasheets

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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: SCF/c-kit Ligand

Stem cell factor (SCF), also known as c-kit ligand (KL), mast cell growth factor (MGF), and steel factor (SLF), is a widely expressed 28 - 40 kDa type I transmembrane glycoprotein. It promotes the survival, differentiation, and mobilization of multiple cell types including myeloid, erythroid, megakaryocytic, lymphoid, germ cell, and melanocyte progenitors. SCF is a primary growth and activation factor for mast cells and eosinophils. Mature human SCF consists of a 189 amino acid (aa) extracellular domain (ECD), a 23 aa transmembrane segment, and a 36 aa cytoplasmic tail. The ECD shows both N-linked and O-linked glycosylation. Proteolytic cleavage at two alternate sites in the extracellular juxtamembrane region releases a 25 kDa soluble molecule which is comparable to the only form produced by Steel-dickie mutant mice. An alternately spliced isoform of human SCF lacks 28 aa that encompasses the primary proteolytic recognition site. Within the ECD of the short isoform (corresponding to this recombinant protein), human SCF shares 75 - 83% aa sequence identity with canine, feline, mouse, and rat SCF. Rat SCF is active on mouse and human cells, but human SCF is only weakly active on mouse cells. Noncovalent dimers of transmembrane or soluble SCF interact with the receptor tyrosine kinase SCF-R/c-kit to trigger receptor dimerization and signaling. SCF assists in the recovery of cardiac function following myocardial infarction by increasing the number of cardiomyocytes and vascular channels.

Long Name:
Stem Cell Factor
Entrez Gene IDs:
4254 (Human); 17311 (Mouse); 60427 (Rat); 403507 (Canine); 493937 (Feline)
Alternate Names:
c-kit Ligand; DCUA; DFNA69; DKFZp686F2250; familial progressive hyperpigmentation 2; FPH2; FPHH; KIT ligand; Kitl; KITLG; KL-1; Mast cell growth factor; MGF; MGFSHEP7; SCF; SCFStem cell factor; SFc-Kit ligand; SHEP7; SLF; steel factor
&#9888; 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. 100 µL Assay Diluent
  4.   Add 100 µL of Assay Diluent to each well.

  5. 100 µL Standard, Control, or Sample
  6.   Add 100 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate each well and wash, repeating the process twice for a total of 3 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  10.   Aspirate and wash 3 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 20 minutes. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   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 for Human SCF Quantikine ELISA Kit

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.

25 Citations: Showing 1 - 10
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  1. KITLG Promotes Glomerular Endothelial Cell Injury in Diabetic Nephropathy by an Autocrine Effect
    Authors: JC Huang, SC Chen, WA Chang, WW Hung, PH Wu, LY Wu, JM Chang, YL Hsu, YC Tsai
    International Journal of Molecular Sciences, 2022-10-03;23(19):.
    Species: Human, Mouse
    Sample Types: Cell Culture Supernates
  2. HIV-1 Tat protein inhibits the hematopoietic support function of human bone marrow mesenchymal stem cells
    Authors: Y Yuan, C Zhou, Q Yang, S Ma, X Wang, X Guo, Y Ding, J Tang, Y Zeng, D Li
    Virus Res., 2019-09-12;273(0):197756.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. Multilevel defects in the hematopoietic niche in essential thrombocythemia
    Authors: T Sun, M Ju, X Dai, H Dong, W Gu, Y Gao, R Fu, X Liu, Y Huang, W Liu, Y Chi, W Wang, H Li, Y Zhou, L Shi, R Yang, L Zhang
    Haematologica, 2019-07-09;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. Development of a Cytocompatible Scaffold from Pig Immature Testicular Tissue Allowing Human Sertoli Cell Attachment, Proliferation and Functionality
    Authors: M Vermeulen, F Del Vento, F de Michele, J Poels, C Wyns
    Int J Mol Sci, 2018-01-12;19(1):.
    Species: Human
    Sample Types: Cell Culture Supernates
  5. Engraftment of chronic myelomonocytic leukemia cells in immunocompromised mice supports disease dependency on cytokines
    Authors: Y Zhang, L He, D Selimoglu-, C Jego, M Morabito, C Willekens, MK Diop, P Gonin, V Lapierre, N Droin, E Solary, F Louache
    Blood Adv, 2017-06-13;1(14):972-979.
    Species: Human
    Sample Types: Plasma
  6. Alteration Analysis of Bone Marrow Mesenchymal Stromal Cells from De Novo Acute Myeloid Leukemia Patients at Diagnosis
    Authors: L Desbourdes, J Javary, T Charbonnie, N Ishac, J Bourgeais, A Iltis, JC Chomel, A Turhan, F Guilloton, K Tarte, MV Demattei, E Ducrocq, F Rouleux-Bo, E Gyan, O Hérault, J Domenech
    Stem Cells Dev., 2017-04-10;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Novel Palladium(II) Complexes that Influence Prominin-1/CD133 Expression and Stem Cell Factor Release in Tumor Cells
    Authors: E Fischer-Fo, R Mikláš, L Rišia?ová, M Cenariu, IG Grosu, P Virag, MP Schrepler, C Tomuleasa, IB Neagoe, F Devínsky, N Miklášová
    Molecules, 2017-03-30;22(4):.
    Species: Human
    Sample Types: Cell Culture Supernates
  8. Angiogenic and vasculogenic factors in the vitreous from patients with proliferative diabetic retinopathy.
    Authors: Abu El-Asrar, Ahmed M, Nawaz, Mohd Imt, Kangave, Dustan, Mairaj Siddiquei, Mohammed, Geboes, Karel
    J Diabetes Res, 2013-03-10;0(0):539658.
    Species: Human
    Sample Types: Vitreous Humor
  9. Mechanism-related circulating proteins as biomarkers for clinical outcome in patients with unresectable hepatocellular carcinoma receiving sunitinib.
    Authors: Harmon CS, DePrimo SE, Raymond E, Cheng AL, Boucher E, Douillard JY, Lim HY, Kim JS, Lechuga MJ, Lanzalone S, Lin X, Faivre S
    J Transl Med, 2011-07-25;9(0):120.
    Species: Human
    Sample Types: Plasma
  10. The change of cytokines in tear and blood after different pterygium operation.
    Authors: Lee JK, Song YS, Shin JS, Kwon YS, Shin MS, Kim JC
    Cytokine, 2009-12-08;49(2):148-54.
    Species: Human
    Sample Types: Tears
  11. Effects of the menstrual cycle on lung function variables in women with asthma.
    Authors: Farha S, Asosingh K, Laskowski D, Hammel J, Dweik RA, Wiedemann HP, Erzurum SC
    Am. J. Respir. Crit. Care Med., 2009-06-11;180(4):304-10.
    Species: Human
    Sample Types: Serum
  12. HLA-DR+ leukocytes acquire CD1 antigens in embryonic and fetal human skin and contain functional antigen-presenting cells.
    Authors: Schuster C, Vaculik C, Fiala C, Meindl S, Brandt O, Imhof M, Stingl G, Eppel W, Elbe-Burger A
    J. Exp. Med., 2009-01-12;206(1):169-81.
    Species: Human
    Sample Types: Cell Culture Supernates
  13. Endothelial progenitor cell levels in obese men with the metabolic syndrome and the effect of simvastatin monotherapy vs. simvastatin/ezetimibe combination therapy.
    Authors: Westerweel PE, Visseren FL, Hajer GR, Olijhoek JK, Hoefer IE, de Bree P, Rafii S, Doevendans PA, Verhaar MC
    Eur. Heart J., 2008-09-28;29(22):2808-17.
    Species: Human
    Sample Types: Serum
  14. Human airway smooth muscle promotes human lung mast cell survival, proliferation, and constitutive activation: cooperative roles for CADM1, stem cell factor, and IL-6.
    Authors: Hollins F, Kaur D, Yang W, Cruse G, Saunders R, Sutcliffe A, Berger P, Ito A, Brightling CE, Bradding P
    J. Immunol., 2008-08-15;181(4):2772-80.
    Species: Human
    Sample Types: Cell Culture Supernates
  15. Isolation and characterization of CD146+ multipotent mesenchymal stromal cells.
    Authors: Sorrentino A, Ferracin M, Castelli G, Biffoni M, Tomaselli G, Baiocchi M, Fatica A, Negrini M, Peschle C, Valtieri M
    Exp. Hematol., 2008-05-27;36(8):1035-46.
    Species: Human
    Sample Types: Cell Culture Supernates
  16. Hypoxia-inducible factor (HIF)-1 alpha directly enhances the transcriptional activity of stem cell factor (SCF) in response to hypoxia and epidermal growth factor (EGF).
    Authors: Han ZB, Ren H, Zhao H, Chi Y, Chen K, Zhou B, Liu YJ, Zhang L, Xu B, Liu B, Yang R, Han ZC
    Carcinogenesis, 2008-03-13;29(10):1853-61.
    Species: Human
    Sample Types: Cell Culture Supernates
  17. Systemic responses after bronchial aspirin challenge in sensitive patients with asthma.
    Authors: Makowska JS, Grzegorczyk J, Bienkiewicz B, Wozniak M, Kowalski ML
    J. Allergy Clin. Immunol., 2007-12-20;121(2):348-54.
    Species: Human
    Sample Types: Serum
  18. C-KIT, by interacting with the membrane-bound ligand, recruits endothelial progenitor cells to inflamed endothelium.
    Authors: Dentelli P, Rosso A, Balsamo A, Colmenares Benedetto S, Zeoli A, Pegoraro M, Camussi G, Pegoraro L, Brizzi MF
    Blood, 2007-02-08;109(10):4264-71.
    Species: Human
    Sample Types: Cell Culture Supernates
  19. Microvascular endothelial cells increase proliferation and inhibit apoptosis of native human acute myelogenous leukemia blasts.
    Authors: Hatfield K, Ryningen A, Corbascio M, Bruserud O
    Int. J. Cancer, 2006-11-15;119(10):2313-21.
    Species: Human
    Sample Types: Cell Culture Supernates
  20. Single administration of stem cell factor, FLT-3 ligand, megakaryocyte growth and development factor, and interleukin-3 in combination soon after irradiation prevents nonhuman primates from myelosuppression: long-term follow-up of hematopoiesis.
    Authors: Drouet M, Mourcin F, Grenier N, Leroux V, Denis J, Mayol JF, Thullier P, Lataillade JJ, Herodin F
    Blood, 2003-10-02;103(3):878-85.
    Species: Primate - Macaca fascicularis (Crab-eating Monkey or Cynomolgus Macaque)
    Sample Types: Plasma
  21. Spontaneous circulation of myeloid-lymphoid-initiating cells and SCID-repopulating cells in sickle cell crisis.
    Authors: Lamming CE, 175, Augustin L, Blackstad M, Lund TC, Hebbel RP, Verfaillie CM
    2003-03-01;111(6):811-9.
    Species: Human
    Sample Types: Serum
  22. Fibroblasts from human spleen regulate NK cell differentiation from blood CD34(+) progenitors via cell surface IL-15.
    Authors: Briard D, Brouty-Boye D, Azzarone B, Jasmin C
    J. Immunol., 2002-05-01;168(9):4326-32.
    Species: Human
    Sample Types: Cell Culture Supernates
  23. Human T-cell leukemia virus type 2 induces survival and proliferation of CD34(+) TF-1 cells through activation of STAT1 and STAT5 by secretion of interferon-gamma and granulocyte macrophage-colony-stimulating factor.
    Authors: Bovolenta C, Pilotti E, Mauri M, Turci M, Ciancianaini P, Fisicaro P, Bertazzoni U, Poli G, Casoli C
    Blood, 2002-01-01;99(1):224-31.
    Species: Human
    Sample Types: Cell Culture Supernates
  24. Human peripheral blood eosinophils express stem cell factor.
    Authors: Hartman M, Piliponsky AM, Temkin V
    Blood, 2001-02-15;97(4):1086-91.
    Species: Human
    Sample Types: Cell Culture Supernates
  25. Stem cell factor (SCF) concentrations in peritoneal fluid of women with or without endometriosis.
    Authors: Osuga Y, Koga K, Tsutsumi O, Igarashi T, Okagaki R, Takai Y, Matsumi H, Hiroi H, Fujiwara T, Momoeda M, Yano T, Taketani Y
    Am. J. Reprod. Immunol., 2000-10-01;44(4):231-5.
    Species: Human
    Sample Types: Serum

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