Human Phospho-M-CSF R DuoSet IC ELISA, 2 Plate

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.

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).

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.

• Reactivity: Hu
• Applications: ELISA
• Conjugate: Biotin

Human Phospho-M-CSF R DuoSet IC ELISA, 2 Plate Summary

• Source: N/A
• Assay Type: Solid Phase Sandwich ELISA
• Inter-Assay: See PDF Datasheet for details
• Intra-Assay: See PDF Datasheet for details
• Spike Recovery: See PDF Datasheet for details
• Sample Volume: See PDF Datasheet for details
• Gene: CSF1R

Applications/Dilutions

• Dilutions:
  • ELISA
• Application Notes: No significant interference observed with available related molecules.

Packaging, Storage & Formulations

• Storage: Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Human Phospho-M-CSF R DuoSet IC ELISA, 2 Plate

• CD115 antigen
• CD115
• c-fms
• colony stimulating factor 1 receptor
• CSF1R
• CSF-1-R
• CSFR
• EC 2.7.10.1
• 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

Background

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.

Publications for M-CSFR/CD115 (DYC3268-2)(1)

Publication using DYC3268-2

• Uitdehaag J, Sunnen C, van Doornmalen A, de Rouw N, Oubrie A, Azevedo R, Ziebell M, Nickbarg E, Karstens W, Ruygrok S Multidimensional profiling of CSF1R screening hits and inhibitors: assessing cellular activity, target residence time, and selectivity in a higher throughput way. J Biomol Screen, 2011-08-26;16(9):1007-17. 2011-08-26 [PMID: 21873591] (Human)

FAQs for M-CSFR/CD115 (DYC3268-2). (Showing 1 - 1 of 1 FAQ).

1. What is the difference between the Quantikine and Duoset ELISA kits?
   • Our Quantikine kits are fully optimized and validated for the sample types listed on the product specific webpage and datasheet, as this can vary between kits. Each kit is supplied ready to use with one pre-coated 96-well plate, a detection antibody directly conjugated to HRP, and all other necessary reagents. These kits are ideal for researchers who want the convenience of a ready to use and optimized ELISA product. Some of these kits are also available prepackaged in larger 6 and 50 plate sizes.Our DuoSet Kits, in contrast, are ELISA development kits containing the capture and detection antibody, the mass-value calibrated standard, and streptavidin-HRP to prepare approximately 5 or 15 plates. Ancillary reagents will need to be used/purchased, and for most kits, we will recommend one of our Ancillary Reagent Kits, which contain the reagents we use ourselves in-house. DuoSet kits are validated only for cell culture supernatant samples and therefore require further development and validation for accurate measurement in more complex samples such as serum and plasma. Our DuoSet Kits offer an economical, flexible alternative for the experienced ELISA user.

Bioinformatics

• Gene Symbol: CSF1R