Clone:
REAL272
Type of antibody:
Releasable antibodies, Primary antibodies, Recombinant antibodies
Applications:
FC
Alternative names:
Csfmr, Fim-2, FMS, M-CSFR, CSF-1R, c-fms

Extended validation for CD115 Antibody, anti-mouse,
REAlease
®

Specificity

Epitope competition
In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.
Other clonesOverlap in epitope recognition with REAL272
AFS98+
T38-320-
460015-
REA827++
Cells were incubated with an excess of purified unconjugated CD115 (REAL272) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison
Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.
View details
Flow cytometric comparison of different clones for CD115. C57BL6/6 splenocytes were stained with CD115 antibodies and with a suitable counterstaining. As a control, CD115 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD115. C57BL6/6 splenocytes were stained with CD115 antibodies and with a suitable counterstaining. As a control, CD115 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD115. C57BL6/6 splenocytes were stained with CD115 antibodies and with a suitable counterstaining. As a control, CD115 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD115. C57BL6/6 splenocytes were stained with CD115 antibodies and with a suitable counterstaining. As a control, CD115 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD115. C57BL6/6 splenocytes were stained with CD115 antibodies and with a suitable counterstaining. As a control, CD115 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Flow cytometric comparison of different clones for CD115. C57BL6/6 splenocytes were stained with CD115 antibodies and with a suitable counterstaining. As a control, CD115 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Fixation data
To provide an indication on how an antibody performs after fixation of cells, in-house data on staining results before and after fixation with 3.7% formaldehyde using Miltenyi Biotec antibodies are provided. Different experimental settings may lead to different results.
No fixation
Post fixation
View details
Comparison of staining pattern on non-fixed and fixed cells using CD115 (REAL272). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
View details
Comparison of staining pattern on non-fixed and fixed cells using CD115 (REAL272). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
Comparison of staining pattern on non-fixed and fixed cells using CD115 (REAL272). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Specifications for CD115 Antibody, anti-mouse,
REAlease
®

Overview

Clone REAL272 is an antibody fragment derived from the full CD115 antibody molecule. It displays no binding to Fc receptors. The recombinantly engineered antibody fragments are multimerized to form the REAlease Complex to bind markers with high avidity.
Clone REAL272 recognizes the protein tyrosine kinase transmembrane receptor for macrophage colony-stimulating factor (M-CSF), also known as CSF-1 and interleukin 34 (IL-34). CD115 is a single-pass type I membrane protein. Receptor activation induces homo-dimerization, which leads to phosphorylation and ubiquitinylation of intracellular residues. CD115 is expressed on mouse monocytes, macrophages, and osteoclasts, as well as on common dendritic cell precursors and macrophage/dendritic cell precursors.
The REAlease Kits consist of the respective fluorochrome-conjugated REAlease Complexes and the REAlease Support Kit for removal of the REAlease Complexes and optional relabeling with different fluorochrome-conjugated REAlease Complexes.

Alternative names

Csfmr, Fim-2, FMS, M-CSFR, CSF-1R, c-fms

Detailed product information

Technical specifications

CloneREAL272
Clonalitymonoclonal
Isotype controlControl Antibody
Hostcell line
Type of antibodyReleasable antibodies, Primary antibodies, Recombinant antibodies
Speciesmouse
AntigenCD115
Alternative names of antigenCsfmr, Fim-2, FMS, M-CSFR, CSF-1R, c-fms
Distribution of antigenmacrophages, monocytes, osteoclasts, dendritic cells
RRIDAB_2811311, AB_2783913, AB_2751450

References for CD115 Antibody, anti-mouse,
REAlease
®

Publications

  1. Sudo, T. et al. (1995) Functional hierarchy of c-kit and c-fms in intramarrow production of CFU-M. Oncogene 11: 2469-2476
  2. Murayama, T. et al. (1999) Intraperitoneal administration of anti-c-fms monoclonal antibody prevents initial events of atherogenesis but does not reduce the size of advanced lesions in apolipoprotein E-deficient mice. Circulation 99: 1740-1746
  3. Sunderkötter, C. et al. (2004) Subpopulations of mouse blood monocytes differ in maturation stage and inflammatory response. J. Immunol. 172: 4410-4417
  4. Auffray, C. et al. (2009)
    CX
    3
    CR1
    +
    CD115
    +
    CD135
    +
    common macrophage/DC precursors and the role of CX
    3
    CR1 in their response to inflammation.
    J. Exp. Med. 206(3): 595-606
  5. Breslin W. L. et al. (2013) Mouse blood monocytes: Standardizing their identification and analysis using CD115. J. Immunol. Methods 390(1-2): 1-8
  6. Lösslein, A.K. et al. (2021) Monocyte progenitors give rise to multinucleated giant cells. Nat Commun. 12(1): 2027

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REAlease
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