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Human M-CSF DuoSet ELISA

Catalog #: DY216
18 Citations 2 Reviews Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
DY216
Ancillary Products Available
DY008B: DuoSet ELISA Ancillary Reagent Kit 2
DY999B: Substrate Reagent Pack
DY999: Substrate Reagent Pack
Human M-CSF ELISA Standard Curve
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Product Details
Procedure
Citations (18)
FAQs
Supplemental Products
Reviews (2)
Summary Product Features Kit Content Other Reagents Required Data Examples Product Datasheets Preparation and Storage Background

Human M-CSF DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
15.6 - 1,000 pg/mL
Sufficient Materials
For fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human M-CSF. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

 

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Scientific Data

N/A Human M-CSF ELISA Standard Curve View Larger

Human M-CSF ELISA Standard Curve

Product Datasheets

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Product Datasheet
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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.
Stability & 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

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

 

Citations for Human M-CSF DuoSet ELISA

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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  1. Signaling events at TMEM doorways provide potential targets for inhibiting breast cancer dissemination
    Authors: Surve, CR;Duran, CL;Ye, X;Chen, X;Lin, Y;Harney, AS;Wang, Y;Sharma, VP;Stanley, ER;McAuliffe, JC;Entenberg, D;Oktay, MH;Condeelis, JS;
    bioRxiv : the preprint server for biology
    Species: Mouse
    Sample Types: Cell Culture Supernates
  2. A Reconstructed Human Melanoma-in-Skin Model to Study Immune Modulatory and Angiogenic Mechanisms Facilitating Initial Melanoma Growth and Invasion
    Authors: Michielon, E;López González, M;Stolk, DA;Stolwijk, JGC;Roffel, S;Waaijman, T;Lougheed, SM;de Gruijl, TD;Gibbs, S;
    Cancers
    Species: Human
    Sample Types: Cell Culture Supernates
  3. CAR T-Cell Targeting of Macrophage Colony-Stimulating Factor Receptor
    Authors: DY Achkova, RE Beatson, J Maher
    Cells, 2022-07-13;11(14):.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. Prediction of emergency cerclage outcomes in women with cervical insufficiency: The role of inflammatory, angiogenic, and extracellular matrix-related proteins in amniotic fluid
    Authors: KN Lee, KH Park, YM Kim, I Cho, TE Kim
    PLoS ONE, 2022-05-10;17(5):e0268291.
    Species: Human
    Sample Types: Amniotic Fluid
  5. Circulating Interleukin-6 and CD16 positive monocytes increase following angioplasty of an arteriovenous fistula
    Authors: S Hakki, EJ Robinson, MG Robson
    Scientific Reports, 2022-01-26;12(1):1427.
    Species: Human
    Sample Types: Plasma
  6. Microfluidic Single-Cell Proteomics Assay Chip: Lung Cancer Cell Line Case Study
    Authors: Y Jung, M Son, YR Nam, J Choi, JR Heath, S Yang
    Micromachines, 2021-09-23;12(10):.
    Species: Human
    Sample Types: Cell Lysates
  7. 6-Shogaol Suppresses 2-Amino-1-Methyl-6-Phenylimidazo [4,5-b] Pyridine (PhIP)-Induced Human 786-O Renal Cell Carcinoma Osteoclastogenic Activity and Metastatic Potential
    Authors: IJ Yeh, SC Chen, MC Yen, YH Wu, CH Hung, PL Kuo
    Nutrients, 2019-09-28;11(10):.
    Species: Human
    Sample Types: Cell Culture Supernates
  8. Immune and Inflammatory Proteins in Cord Blood as Predictive Biomarkers of Retinopathy of Prematurity in Preterm Infants
    Authors: YJ Park, SJ Woo, YM Kim, S Hong, YE Lee, KH Park
    Invest. Ophthalmol. Vis. Sci., 2019-09-03;60(12):3813-3820.
    Species: Human
    Sample Types: Plasma
  9. The human glomerular endothelial cells are potent pro-inflammatory contributors in an in vitro model of lupus nephritis
    Authors: P Dimou, RD Wright, KL Budge, A Midgley, SC Satchell, M Peak, MW Beresford
    Sci Rep, 2019-06-06;9(1):8348.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. Colony?stimulating factor 1 receptor inhibition blocks macrophage infiltration and endometrial cancer cell proliferation
    Authors: F Hua, Y Tian, Y Gao, C Li, X Liu
    Mol Med Rep, 2019-02-18;0(0):.
    Species: Human
    Sample Types: Tissue Homogenates
  11. The RUNX1/IL-34/CSF-1R axis is an autocrinally regulated modulator of resistance to BRAF-V600E inhibition in melanoma
    Authors: O Giricz, Y Mo, KB Dahlman, XM Cotto-Rios, C Vardabasso, H Nguyen, B Matusow, M Bartenstei, V Polishchuk, DB Johnson, TD Bhagat, R Shellooe, E Burton, J Tsai, C Zhang, G Habets, JM Greally, Y Yu, PA Kenny, GB Fields, K Pradhan, ER Stanley, E Bernstein, G Bollag, E Gavathioti, BL West, JA Sosman, AK Verma
    JCI Insight, 2018-07-26;3(14):.
    Species: Human
    Sample Types: Cell Culture Supernates
  12. Novel GM-CSF signals via IFN-?R/IRF-1 and AKT/mTOR license monocytes for suppressor function
    Authors: E Ribechini, JA Hutchinson, S Hergovits, M Heuer, J Lucas, U Schleicher, AL Jordán Gar, SJ Potter, P Riquelme, H Brackmann, N Müller, H Raifer, I Berberich, M Huber, A Beilhack, M Lohoff, C Bogdan, M Eyrich, HM Hermanns, EK Geissler, MB Lutz
    Blood Adv, 2017-06-07;1(14):947-960.
    Species: Human
    Sample Types: Plasma
  13. Glioblastoma-derived Macrophage Colony-stimulating Factor (MCSF) Induces Microglial Release of Insulin-like Growth Factor-binding Protein 1 (IGFBP1) to Promote Angiogenesis.
    Authors: Nijaguna M, Patil V, Urbach S, Shwetha S, Sravani K, Hegde A, Chandramouli B, Arivazhagan A, Marin P, Santosh V, Somasundaram K
    J Biol Chem, 2015-08-05;290(38):23401-15.
    Species: Human
    Sample Types: Cell Culture Supernates
  14. CD137 ligand signalling induces differentiation of primary acute myeloid leukaemia cells.
    Authors: Cheng K, Wong S, Linn Y, Ho L, Chng W, Schwarz H
    Br J Haematol, 2014-01-15;165(1):134-44.
    Species: Human
    Sample Types: Cell Culture Supernates
  15. Derepression of an endogenous long terminal repeat activates the CSF1R proto-oncogene in human lymphoma.
    Authors: Lamprecht B, Walter K, Kreher S, Kumar R, Hummel M, Lenze D, Kochert K, Bouhlel MA, Richter J, Soler E, Stadhouders R, Johrens K, Wurster KD, Callen DF, Harte MF, Giefing M, Barlow R, Stein H, Anagnostopoulos I, Janz M, Cockerill PN, Siebert R, Dorken B, Bonifer C, Mathas S
    Nat. Med., 2010-05-02;16(5):571-9.
    Species: Human
    Sample Types: Cell Culture Supernates
  16. Epigenetic control of MHC class II expression in tumor-associated macrophages by decoy receptor 3.
    Authors: Chang YC, Chen TC, Lee CT, Yang CY, Wang HW, Wang CC, Hsieh SL
    Blood, 2008-03-18;111(10):5054-63.
    Species: Human
    Sample Types: Whole Cells
  17. 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, 2007-09-11;110(13):4319-30.
    Species: Human
    Sample Types: Cell Culture Supernates
  18. Ligation of the FcR gamma chain-associated human osteoclast-associated receptor enhances the proinflammatory responses of human monocytes and neutrophils.
    Authors: Merck E, Gaillard C, Scuiller M, Scapini P, Cassatella MA, Trinchieri G, Bates EE
    J. Immunol., 2006-03-01;176(5):3149-56.
    Species: Human
    Sample Types: Cell Culture Supernates

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Reviews for Human M-CSF DuoSet ELISA

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