Human Phospho-M-CSF R DuoSet IC ELISA

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Figure 1.  The Human Phospho-M-CSF Receptor DuoSet IC ELISA is more sensitive than immunoprecipitation (IP)-Western analysis
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Product Details
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Human Phospho-M-CSF R DuoSet IC ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
96-well strip plate
Sample Volume Required
Cell lysates (100 µL)
Sufficient Materials
Kits available for two, five, or fifteen 96-well plates*
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 IC ELISA contains the basic components required for the development of sandwich ELISAs to measure tyrosine-phosphorylated human M-CSF R / CD115 in cell lysates. An immobilized capture antibody specific for M-CSF R / CD115 binds both phosphorylated and unphosphorylated M-CSF R / CD115. After washing away unbound material, a biotinylated detection antibody is used to detect only phosphorylated receptor, utilizing a standard HRP format.

Product Features

  • Optimized capture and detection antibody pairings and recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Available in 2, 5, and 15-(96-well) plate pack sizes
  • Economical alternative to Western blot

Kit Content

  • Capture Antibody
  • Conjugated Detection Antibody
  • Calibrated Immunoassay Standard or Control

Other Reagents Required

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2O4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or equivalent

Lysis Buffer*

IC Diluent*

Blocking Buffer*

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), or equivalent

Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent

*For the Lysis Buffer, IC Diluent, and Blocking BUffer recommended for a specific DuoSet ELISA Development Kit, please see the product

Scientific Data

Figure 1. The Human Phospho-M-CSF Receptor DuoSet IC ELISA is more sensitive than immunoprecipitation (IP)-Western analysis The human acute monocytic leukemia cell line THP-1 was treated with 600 ng/mL recombinant human M-CSF (R&D Systems, Catalog #216-MC) for five minutes to induce tyrosine phosphorylation of M-CSF Receptor. Lysates were serially diluted and analyzed by (A) IP-Western blot and (B) this DuoSet IC ELISA. IPs were done using an anti-M-CSF Receptor polyclonal antibody and protein G agarose. Immunoblots were incubated with a biotinylated anti-phosphotyrosine monoclonal antibody (R&D Systems, Catalog # BAM1676) to detect phospho-M-CSF Receptor. Bands were visualized with Streptavidin-HRP (R&D Systems, Cat # DY998) followed by chemiluminescent detection using WesternGloTM Chemiluminescent Detection Substrate (R&D Systems, Catalog # AR004). Human Phospho-M-CSF Receptor can be detected in this DuoSet IC ELISA by using approximately 4 to 8 times less lysate than is needed for a conventional IP-Western blot.

Figure 2. The Human Phospho-M-CSF Receptor DuoSet IC ELISA detects ligand-induced Flt-3 tyrosine phosphorylation THP-1 cells were untreated or treated with 600 ng/mL recombinant human M-CSF for five minutes. ELISA and IP-Western blot (inset) analyses were done using 100 μg and 400 μg of lysate, respectively. IP-Western blots for phospho-M-CSF Receptor (p-M-CSF R) were done as described in Figure 1. Blots were stripped and total M-CSF Receptor was detected using a biotinylated anti-M-CSF Receptor polyclonal antibody (R&D Systems, Catalog #BAF329).

Figure 3. The specificity of the Human Phospho-M-CSF ReceptorDuoSet IC ELISA is confirmed by receptor competition beta THP-1 cells were treated with 600 ng/mL recombinant human M-CSF for five minutes. The indicated amounts of recombinant extracellular domains of human M-CSF Receptor (R&D Systems, Catalog #329-MR), human SCF sR/c-kit (R&D Systems, Catalog #332-SR), human Flt-3 (R&D Systems, Catalog# 368-ST), human PDGF sR alpha (R&D Systems, Catalog #322-PR), or human PDGF R beta (R&D Systems, Catalog #385-PR) were added to 100 μg lysate and analyzed using this DuoSet IC ELISA. Competition was observed only with recombinant Human M-CSF Receptor.

Product Datasheets

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Preparation and Storage

The product is shipped with polar packs. 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 R/CD115

M-CSF R/CD115, the product of the c-fms proto-oncogene, is a member of the type III subfamily of receptor tyrosine kinases that also includes c-kit, the receptor for SCF, and the alpha and beta receptors for PDGF. M-CSF R/CD115 is expressed primarily on cells of the monocyte/macrophage lineage and in various tissues of the developing placenta.

Long Name:
Macrophage Colony Stimulating Factor Receptor
Entrez Gene IDs:
1436 (Human); 12978 (Mouse)
Alternate Names:
CD115 antigen; CD115; c-fms; colony stimulating factor 1 receptor; CSF1R; CSF-1-R; CSFR; EC; FMS proto-oncogene; FMSFIM2; macrophage colony stimulating factor I receptor; macrophage colony-stimulating factor 1 receptor; McDonough feline sarcoma viral (v-fms) oncogene homolog; M-CSF R; MCSFR; M-CSFR; Proto-oncogene c-Fms

Citation for Human Phospho-M-CSF R DuoSet IC 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.

1 Citation: Showing 1 - 1

  1. Multidimensional profiling of CSF1R screening hits and inhibitors: assessing cellular activity, target residence time, and selectivity in a higher throughput way.
    Authors: Uitdehaag J, Sunnen C, van Doornmalen A, de Rouw N, Oubrie A, Azevedo R, Ziebell M, Nickbarg E, Karstens W, Ruygrok S
    J Biomol Screen, 2011-08-26;16(9):1007-17.
    Species: Human
    Sample Types: Cell Lysates


  1. Which phosphorylated sites are recognized by this assay?

    • This assay utilizes an anti-phosphorylated tyrosine monoclonal detection antibody, and it recognizes all phosphorylated tyrosine residues.

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