- Predicted Species Reactivity:
- Product Format:
- Purified IgG - liquid
- IHC-AFF, FC, IP
- Additional Information:
- Fusion Partners: Spleen cells from an immunised Wistar rat were fused with cells of the SP2/0-Ag14 mouse myeloma cell line
- Preparation: Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
Preservative Stabilisers: 0.09% - Sodium Azide
- Approx Protein Conc: IgG concentration 0.5 mg/ml
Buffer Solutions: TRIS buffered saline
- Reconstitution and Storage:
- Store at +4oC or at -20oC if preferred.
This product should be stored undiluted.
Storage in frost-free freezers is not recommended. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
- NCBI Gene Id:
- Protein Name:
- Receptor-type tyrosine-protein phosphatase C
- Alias Symbols:
- loc, B220, Cd45, Ly-5, T200, CD45R, Lyt-4, Ptprc
- Replacement Item:
- This antibody may replace item sc-1123 from Santa Cruz Biotechnology.
- Description of Target:
- RAT ANTI MOUSE CD45
- Protein Size (# AA):
- Tissue Tool:
- Find tissues and cell lines supported by DNA array analysis to express Ptprc.
- RNA Seq:
- Find tissues and cell lines supported by RNA-seq analysis to express Ptprc.
- The immunogen for anti-Ptprc antibody: purified B cells from mouse lymph nodes
- Predicted Homology Based on Immunogen Sequence:
- Printable datasheet for anti-Ptprc antibody - OASA05279
- Application Info:
- Flow Cytometry: 1/50 - 1/100
Immunohistology - Frozen: 1/50 - 1/100
- Protocol Information:
- Citation: 1: Copland DA, Calder CJ, Raveney BJ, Nicholson LB, Phillips J, Cherwinski H, Jenmalm M, Sedgwick JD, Dick AD. Monoclonal antibody-mediated CD200 receptor signaling suppresses macrophage activation and tissue damage in experimental autoimmune uveoretinitis. Am J Pathol. 2007 Aug;171(2):580-8. Epub 2007 Jun 28. PubMed PMID: 17600119; PubMed Central PMCID: PMC1934542.
Experiment Name: 1. EAU Induction and Scoring2. Isolation of Retinal Myeloid Cells
Experiment Background: 1. In this study, David et al. show that inducing myeloid CD200R signaling via the systemic administration of an agonist rat anti-mouse mAb, DX109 successfully suppresses an archetypal organ-specific T-cell-mediated disease, EAU.2. A model of CD4+ T-cell organ-specific autoimmunity, experimental autoimmune uveoretinitis (EAU) was used in these studies because of extensive neuronal CD200 expression in the eye where tissue destruction is largely mediated via activated macrophages and NO damage. Systemic administration of a rat anti-mouse CD200R monoclonal antibody (mAb) (DX109) resulted in suppression of EAU and tissue damage even in the presence of systemic T-cell proliferation and IFNg production.
Experimental Steps: 1. EAU Induction and Scoring: C57BL/6 and CD200-/- mice were immunized subcutaneously in one flank with 500 µg/mouse hRBP-3 1-20 peptide in phosphate (PBS) (2% dimethyl sulfoxide), in emulsion with complete Freud’s adjuvant [1 mg/ml; 1:1 (v/v)] supplemented with 1.5 mg/ml Mycobacterium tuberculosis complete H37 RA (BD Biosciences) and 1.5 µg of Bordetella pertussis toxin (Sigma-Aldrich, Dorset, UK) intraperitoneally (i.p.). B10.RIII mice were immunized subcutaneously in one flank with 50 µg/mouse IRBP 161-180 peptide in PBS (2% dimethyl sulfoxide), in complete Freud’s adjuvant, with an additional i.p. injection of 1 µg of Bordetella pertussis toxin. At various time points after immunization, eyes were enucleated and carefully snap-frozen, oriented in optimal cutting temperature compound (R. Lamb Ltd., East Sussex, UK). Serial 12-µm sections were thawed at room temperature and fixed in acetone for 10 minutes. Sections were stained with rat anti-mouse monoclonal anti-CD45 antibody (Oxford, UK) and counterstained with hematoxylin (ThermoShandon, Pittsburgh, PA). Sections were scored for inflammatory infiltrate (presence of CD45-positive cells) and structural disease (disruption of morphology).2. Isolation of Retinal Myeloid Cells: Eyes were enucleated from immunized mice, and the retinas of each animal were dissected microscopically in ice-cold 1% bovine serum albumin (Sigma-Aldrich) in PBS (1% PBSA). Retinas were then mechanically disrupted through a 40-µm cell strainer to obtain a singlecell suspension. A sample was removed to determine the purity and percentage of F4/80-positive macrophages by staining cells with fluorescein isothiocyanate-conjugated anti-mouse F4/80 before flow cytometric analysis. Cells were plated at 5 x 105 F4/80-positive cells/ml for 2 hours, to allow the isolation of macrophages from other cell contaminants via plastic adherence. The medium was replaced and assays performed.
Other Reagents Used: Penicillin, streptomycin, L-glutamine, sodium pyruvate, and 2-mercaptoethanol
Number Of Protocols: 2
- Target Reference:
- 1. Clausen, B.H. et al. (2008) Interleukin-1beta and tumor necrosis factor-alpha are expressed by different
subsets of microglia and macrophages after ischemic stroke in mice. J. Neuroinflammation. 5:46.
2. Balogh, P., Bal¡zs, M. and A. Kum¡novicx (1995) Modulatory effect of CD45 on the MHC Class II-induced homotypic aggregation of B cells in mice. In 9th International Congress of Immunology, Abstract book p. 72. No. 422.