- Predicted Species Reactivity:
- Product Format:
- Purified IgG - liquid
- IHC-AFF, FC, IHC-FFPE
- Additional Information:
- Fusion Partners: Spleen cells from immunised BALB/c mice were fused with cells from the NS1 mouse myeloma cell line.
- Preparation: Purified IgG prepared by affinity chromatography on Protein G
Preservative Stabilisers: 0.09% - Sodium Azide
- Approx Protein Conc: IgG concentration 1.0 mg/ml
- 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:
- Lca, RT7, CD45, L-CA, T200, Ptprc
- Replacement Item:
- This antibody may replace item sc-1123 from Santa Cruz Biotechnology.
- Description of Target:
- MOUSE ANTI RAT CD45RC
- 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: pHA stimulated rat lymphocytes
- Predicted Homology Based on Immunogen Sequence:
- Printable datasheet for anti-Ptprc antibody - OASA04013
- Application Info:
- Flow Cytometry: 1/100
Immunohistology - Frozen: 1/100 - 1/1000
Immunohistology - Paraffin(1): 1/100 - 1/1000
(1)Special conditions apply. - See [*].
- Application Data:
- Application #1: Staining of rat spleen cells with MOUSE.ANTI RAT CD45RC:Alexa Fluor 647
Application #2: Staining of rat spleen cells with MOUSE.ANTI RAT CD45RC:FITC
- Protocol Information:
- Citation: 1: Aiello S, Noris M, Piccinini G, Tomasoni S, Casiraghi F, Bonazzola S, Mister M, Sayegh MH, Remuzzi G. Thymic dendritic cells express inducible nitric oxide synthase and generate nitric oxide in response to self- and alloantigens. J Immunol. 2000 May 1;164(9):4649-58. PubMed PMID: 10779769.
Experiment Name: Double Labeling
Experiment Background: Lhymocytes maturing in the thymus undergo clonal deletion/apoptosis when they encounter self- or allo-Ags presented by dendritic cells (DCs). How this occurs is a matter of debate, but NO may play a role given its ability of inducing apoptosis of these cells. Analysis by FACS and by double labeling of cytocentrifuged preparations showed that DCs and MF both express iNOS within APC. Analysis of a purified preparation of DCs confirmed that these cells express high levels of iNOS and produce large amounts of NO in basal conditions. The capacity of DCs to generate NO was enhanced by exposure to rat albumin, a self-protein, and required a fully expressed process of Ag internalization, processing, and presentation. Peptides derived from portions of class II MHC molecules up-regulate iNOS expression and NO production by DCs as well, both in self and allogeneic combinations, suggesting a role of NO in both self and acquired tolerance. et al. also found that NO induced apoptosis of rat double-positive thymocytes, the effect being more evident in anti-CD3-stimulated cells. Altogether, the present findings might suggest that DC-derived NO is at least one of the soluble factors regulating events, in the thymus, that follow recognition of self- and allo-Ags.
Experimental Steps: Thymi (10–20 for each experiment) were cut into small fragments and digested with collagenase and a total thymocyte suspension was obtained.After that the suspension was cultured. The adherent cells comprised two populations: some cells showed the characteristics of DCs and other cells were circumferentially spread, ruffled cells with many vesicles. The adherent cells were cultured. After the overnight culture, the floating cells were collected. Cells that detached after the overnight culture consisted of a large cell population that was MHC I+, MHC II++, CD4-, CD8-/+, CD45RA-, CD3-, and OX62+, consistent with the expected profile of DCs..However, a considerable number were ED2+ and ED3+, suggesting that some cells also detached during the overnight culture. Thus, we called these cells thymic APC.In selected experiments, MΦwere collected, by treating the petri dishes with EDTA 30 mM, and analyzed by FACSThymic APC were centrifuged on 55% Percoll (Pharmacia LKB, Uppsala, Sweden) solution for 20 min at 4C, and the low-density fraction was collected and subjected to two rounds of plastic adherence for 30 min at 37C. The final enrichment for DCs was routinely performed by removing T cells and MΦ using magnetic beads. “Briefly, cells were incubated with a mixture of appropriate dilutions of R73, V65, OX12, OX22, OX33, 341.1, ED2, and ED3 mAbs for 30 min at 4C, washed three times, and then incubated with rat anti-mouse IgG-coated magnetic beads for 30 min at 4C in agitation. “
Other Reagents Used: Dowex AG 50 WX-8HEPESbrefeldin A (BfA)chloroquine (ChlQ)amiloride (AML)
Number Of Protocols: 1
- Target Reference:
- 1. Spickett, G.P. et al. (1983) MRC OX-22, a monoclonal antibody that labels a new subset of T lymphocytes and reacts with the high molecular weight form of the leukocyte-common antigen. J. Exp. Med. 158:795-810.
2. Arthur, R.P. and Mason, D.W. (1986) T cells that help B cell responses to soluble antigen are distinguishable from those producing interleukin 2 on mitogenic or allogeneic stimulation. J. Exp. Med. 163:774-786.
3. Pelegri, C et al. (2001) Prevention of adjuvant arthritis by the W3/25 anti-CD4 monoclonal antibody is associated with a decrease of blood CD4+CD45RChigh T cells. Clin. Exp. Immunol. 125:470-477.