mAbs Against HUMAN Protein
CSF1R
| STATUS: |  |
|---|---|
| CONTACT INFORMATION: | Monoclonal Antibodies Unit, Centro Nacional de Investigaciones Oncológicas |
| STATUS: | Validated |
| TYPE: | mouse anti human |
| CLONE NAME: | FER216 |
| PROTEIN: | Macrophage colony-stimulating factor 1 receptor precursor |
| PROTEIN WEB: | http://www.ncbi.nlm.nih.gov/protein/NP_005202.2 |
| ANTIGEN USED: | ecCSF1R-Fc-6His recombinant protein |
| FUSION PARTNER: | NS1/Ag4-1 (NS1) cells |
| ISOTYPE: | IgG1 |
| SPECIES REACTIVITY: | Human |
| PREPARATION AND STORAGE: | Aliquot and store at 4C. Do not freeze |
Description
The protein encoded by this gene is the receptor for colony stimulating factor 1, a cytokine which controls the production, differentiation, and function of macrophages. This receptor mediates most if not all of the biological effects of this cytokine. Ligand binding activates the receptor kinase through a process of oligomerization and transphosphorylation. The encoded protein is a tyrosine kinase transmembrane receptor and member of the CSF1/PDGF receptor family of tyrosine-protein kinases. [provided by RefSeq, Dec 2013].
Publication describing antibody characterization/validation
Martín-Moreno AM, Roncador G, Maestre L, Mata E, Jiménez S, Martínez-Torrecuadrada JL, Reyes-García AI, Rubio C, Tomás JF, Estévez M, Pulford K, Piris MA, García JF. CSF1R Protein Expression in Reactive Lymphoid Tissues and Lymphoma: Its Relevance in Classical Hodgkin Lymphoma. PLoS One 2015 Jun 12;10(6).
Applications
| IHC Techniques | Clone | Dilution | Antibody concentration | Antigen retrieval method | Visualization kit | +/- control | Protein localization | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|---|
| Frozen tissue and cytospins | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | FER216 | Neat supernatant | Tonsil / | ||||||
| Paraffin tissue | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | FER216 | 1:10 | supernatant | Tonsil / | Available | ||||
| Immunofluorescence | |||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | FER216 | supernatant | Tonsil / | Available | |||||
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- FER216 mAb in transfected cells
- Validation of FER216 monoclonal antibody in HEK293T/CSF1R transfected cells. HEK293T/FGFR1 transfected cells were used as negative control. anti-CSF1R antibody (clone 61710) confirms transfection efficiency.
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- FER216 mAb in human frozen tonsil section
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- FER216 staining in human paraffin embeded tissues
- The range of expression of CSF1R in tisues varied considerably between the different functional tissue compartments. The higher frequency of CSF1R+ macrophages was detected in the paracortex of tonsil.
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- Immunofluorescence of FER216 mAb in human paraffin embeded tonsil.
| WB Techniques | Clone | Dilution | Antibody concentration | +/- control | Expected MW | Observed Mw | Positivity in other species | Protocol |
|---|---|---|---|---|---|---|---|---|
| Western Blotting | ||||||||
Recommended Result obtained is satisfactory. The reagent can be use in this application | FER216 | 1:2 | supernatant | Tonsil / L540 cell line | Available | |||
| Immunoprecipitation | ||||||||
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- FER216 mAb in western blotting
- Lane1 HEK-CSF1R-Fc transfected cells (20ug) (+)
Lane 2 HEK-FGFR1 transfected cells (20ug) (-)
Lane 3 Human tonsil (150ug) (+)
Lane 4 Human testicle (150ug) (-)
Lane 5 Hodgking lymphoma case (150ug) (+)
Lane 6 Hodgking lymphoma case (150ug) (+)
Lane 7 Hodgking lymphoma case (150ug) (+)
Lane 8 L428 cell line (150ug)(-)
Lane 9 L1236 cell line (150ug) (-)
Lane 10 KM-H2 cell line (150ug) (-)
Lane 11 HDLM2 cell line (150ug) (-)
Lane 12 L540 cell line (150ug) (-)
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- Immunoprecipitation with FER216
- Immunoprecipitation of protein extracts from human tonsil, one HD case, KMH2, and L540 with FER216 mAb (30ul/IP) followed by western blotting with FER216 (1:5 dilution).
| FLOW CYTOMETRY | Clone | Dilution | +/- control | Type of fluorocrom | Protocol |
|---|---|---|---|---|---|
Recommended Result obtained is satisfactory. The reagent can be use in this application | FER216 | / | PE |
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- Expression of CSF1R (FER216) on Karpas 299 human cell line.