Mouse M-CSF Quantikine ELISA Kit

  (18 citations)
(2 Reviews)
    
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Assay Procedure
Citations (18)
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  • 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 (13 uL), Serum (13 uL), EDTA Plasma (13 uL)
  • Sensitivity
    5 pg/mL
  • Assay Range
    31.2 - 2,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma)
  • Specificity
    Natural and recombinant mouse M-CSF
  • 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 M-CSF Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse M-CSF levels in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant mouse M-CSF and antibodies raised against the recombinant factor. Results obtained for naturally occurring mouse M-CSF showed linear curves that were parallel to the standard curves obtained using the kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse M-CSF.

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, Serum, EDTA Plasma
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 44 174 558 46 174 554
Standard Deviation 3.9 10.2 38.9 4 10.1 35.2
CV% 8.9 5.9 7 8.7 5.8 6.4

Recovery

The recovery of mouse M-CSF spiked to three levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=5) 104 85-120
EDTA Plasma (n=6) 101 89-112
Serum (n=6) 97 85-115
Linearity
To assess the linearity of the assay, five or more samples containing and/or spiked with various concentrations of mouse M-CSF in each matrix were diluted with Calibrator Diluent and then assayed.
 M-CSF [HRP]
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: M-CSF
M-CSF, also known as CSF-1, is a four-alpha-helical-bundle cytokine that is the primary regulator of macrophage survival, proliferation and differentiation. M-CSF is also essential for the survival and proliferation of osteoclast progenitors. M-CSF also primes and enhances macrophage killing of tumor cells and microorganisms, regulates the release of cytokines and other inflammatory modulators from macrophages, and stimulates pinocytosis. M-CSF increases during pregnancy to support implantation and growth of the decidua and placenta. Sources of M-CSF include fibroblasts, activated macrophages, endometrial secretory epithelium, bone marrow stromal cells and activated endothelial cells. The M-CSF receptor (c-fms) transduces its pleotropic effects and mediates its endocytosis. M-CSF mRNAs of various sizes occur. Full length human M-CSF transcripts encode a 522 amino acid (aa) type I transmembrane (TM) protein with a 464 aa extracellular region, a 21 aa TM domain, and a 37 aa cytoplasmic tail that forms a 140 kDa covalent dimer. Differential processing produces two proteolytically cleaved, secreted dimers. One is an N- and O- glycosylated 86 kDa dimer, while the other is modified by both glycosylation and chondroitin-sulfate proteoglycan (PG) to generate a 200 kDa subunit. Although PG-modified M-CSF can circulate, it may be immobilized by attachment to type V collagen. Shorter transcripts encode M-CSF that lack cleavage and PG sites and produce an N-glycosylated 68 kDa TM dimer and a slowly produced 44 kDa secreted dimer. Although forms may vary in activity and half-life, all contain the N-terminal 150 aa portion that is necessary and sufficient for interaction with the M-CSF receptor. The first 223 aa of mature human M-CSF shares 88%, 86%, 81% and 74% aa identity with corresponding regions of dog, cow, mouse and rat M-CSF, respectively. Human M-CSF is active in the mouse, but mouse M-CSF is reported to be species-specific.
  • Long Name:
    Macrophage Colony Stimulating Factor
  • Entrez Gene IDs:
    1435 (Human); 12977 (Mouse)
  • Alternate Names:
    colony stimulating factor 1 (macrophage); CSF1; CSF-1; Lanimostim; macrophage colony stimulating factor; macrophage colony-stimulating factor 1; MCSF; M-CSF; MCSFlanimostim; MGC31930
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
  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.
  7.   Aspirate each well and wash, repeating the process 4 times for a total of 5 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.
  10.   Aspirate and wash 5 times.

  11. 100 µL Substrate Solution
  12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. 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.
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.

18 Citations: Showing 1 - 10
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Species
Sample Type
  1. Macrophage-derived granulin drives resistance to immune checkpoint inhibition in metastatic pancreatic cancer
    Authors: V Quaranta, C Rainer, SR Nielsen, ML Raymant, MS Ahmed, DD Engle, A Taylor, T Murray, F Campbell, DH Palmer, DA Tuveson, A Mielgo, MC Schmid
    Cancer Res., 2018;0(0):.
    Species: Mouse
    Sample Type: Cell Culture Supernates
  2. Calf Spleen Extractive Injection protects mice against cyclophosphamide-induced hematopoietic injury through G-CSF-mediated JAK2/STAT3 signaling
    Authors: W Lu, D Jia, S An, M Mu, X Qiao, Y Liu, X Li, D Wang
    Sci Rep, 2017;7(1):8402.
    Species: Mouse
    Sample Type: Plasma
  3. iRhom2 regulates CSF1R cell surface expression and non-steady state myelopoiesis in mice
    Eur J Immunol, 2016;0(0):.
    Species: Mouse
    Sample Type: Serum
  4. Monocyte/macrophage lineage commitment and distribution are affected by the lack of regulatory T cells in scurfy mice
    Eur J Immunol, 2016;0(0):.
    Species: Mouse
    Sample Type: Cell Culture Supernates
  5. SHIP1-expressing mesenchymal stem cells regulate hematopoietic stem cell homeostasis and lineage commitment during aging.
    Authors: Iyer S, Brooks R, Gumbleton M, Kerr W
    Stem Cells Dev, 2015;24(9):1073-81.
    Species: Mouse
    Sample Type: Serum
  6. Steady-state neutrophil homeostasis is dependent on TLR4/TRIF signaling.
    Authors: Bugl S, Wirths S, Radsak M, Schild H, Stein P, Andre M, Muller M, Malenke E, Wiesner T, Marklin M, Frick J, Handgretinger R, Rammensee H, Kanz L, Kopp H
    Blood, 2013;121(5):723-33.
    Species: Mouse
    Sample Type: Plasma
  7. KIT oncogene inhibition drives intratumoral macrophage M2 polarization.
    Authors: Cavnar, Michael, Zeng, Shan, Kim, Teresa S, Sorenson, Eric C, Ocuin, Lee M, Balachandran, Vinod P, Seifert, Adrian M, Greer, Jonathan, Popow, Rachel, Crawley, Megan H, Cohen, Noah A, Green, Benjamin, Rossi, Ferdinan, Besmer, Peter, Antonescu, Cristina, DeMatteo, Ronald P
    J Exp Med, 2013;210(13):2873-86.
    Species: Mouse
    Sample Type: Serum
  8. The combination of the histone deacetylase inhibitor vorinostat and synthetic triterpenoids reduces tumorigenesis in mouse models of cancer.
    Authors: Tran, Kim, Risingsong, Renee, Royce, Darlene, Williams, Charlott, Sporn, Michael, Pioli, Patricia, Gediya, Lalji K, Njar, Vincent, Liby, Karen T
    Carcinogenesis, 2013;34(1):199-210.
    Species: Mouse
    Sample Type: Cell Culture Supernates
  9. Osteoclasts are dispensable for hematopoietic stem cell maintenance and mobilization.
    Authors: Miyamoto K, Yoshida S, Kawasumi M
    J. Exp. Med., 2011;208(11):2175-81.
    Species: Mouse
    Sample Type: Serum
  10. Temporal changes in myeloid cells in the cervix during pregnancy and parturition.
    Authors: Timmons BC, Fairhurst AM, Mahendroo MS
    J. Immunol., 2009;182(5):2700-7.
    Species: Mouse
    Sample Type: Tissue Homogenates
  11. A potential role of thymic stromal lymphopoietin in the recruitment of macrophages to mouse intervertebral disc cells via monocyte chemotactic protein 1 induction: implications for herniated discs.
    Authors: Ohba T, Haro H, Ando T, Koyama K, Hatsushika K, Suenaga F, Ohnuma Y, Nakamura Y, Katoh R, Ogawa H, Hamada Y, Nakao A
    Arthritis Rheum., 2008;58(11):3510-9.
    Species: Mouse
    Sample Type: Cell Culture Supernates
  12. Bacterial lipopolysaccharide induces osteoclast formation in RAW 264.7 macrophage cells.
    Authors: Islam S, Hassan F, Tumurkhuu G, Dagvadorj J, Koide N, Naiki Y, Mori I, Yoshida T, Yokochi T
    Biochem. Biophys. Res. Commun., 2007;360(2):346-51.
    Species: Mouse
    Sample Type: Cell Culture Supernates
  13. IL-7 induces myelopoiesis and erythropoiesis.
    Authors: Aiello FB, Keller JR, Klarmann KD, Dranoff G, Mazzucchelli R, Durum SK
    J. Immunol., 2007;178(3):1553-63.
    Species: Mouse
    Sample Type: Cell Culture Supernates
  14. Concerted action of Smad and CREB-binding protein regulates bone morphogenetic protein-2-stimulated osteoblastic colony-stimulating factor-1 expression.
    Authors: Ghosh-Choudhury N, Singha PK, Woodruff K, St Clair P, Bsoul S, Werner SL, Choudhury GG
    J. Biol. Chem., 2006;281(29):20160-70.
    Species: Mouse
    Sample Type: Cell Culture Supernates
  15. Perforin-deficient CD8+ T cells mediate fatal lymphocytic choriomeningitis despite impaired cytokine production.
    Authors: Storm P, Bartholdy C, Sorensen MR, Christensen JP, Thomsen AR
    J. Virol., 2006;80(3):1222-30.
    Species: Mouse
    Sample Type: Serum
  16. Interleukin-17 as a recruitment and survival factor for airway macrophages in allergic airway inflammation.
    Authors: Sergejeva S, Ivanov S, Lotvall J, Linden A
    Am. J. Respir. Cell Mol. Biol., 2005;33(3):248-53.
    Species: Mouse
    Sample Type: BALF
  17. The cell-surface isoform of colony stimulating factor 1 (CSF1) restores but does not completely normalize fecundity in CSF1-deficient mice.
    Authors: Ovadia S, Insogna K, Yao GQ
    Biol. Reprod., 2005;74(2):331-6.
    Species: Mouse
    Sample Type: Tissue Homogenates
  18. Differential roles of CC chemokine ligand 2/monocyte chemotactic protein-1 and CCR2 in the development of T1 immunity.
    Authors: Traynor TR, Herring AC, Dorf ME, Kuziel WA, Toews GB, Huffnagle GB
    J. Immunol., 2002;168(9):4659-66.
    Species: Mouse
    Sample Type: BALF
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Sample Tested Activated mouse CD8 T cell
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