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Purified Rat Anti-Mouse CD210
Purified Rat Anti-Mouse CD210

Flow cytometric analysis of CD210 expression on mouse splenocytes. Mouse B220+ splenocytes were stained with Purified Rat Anti-Mouse CD210 (Cat. No. 559912) followed by Biotin Goat Anti-Rat Ig (Cat. No. 554014) and Streptavidin-PE (Cat. No. 554061) in a three-layer staining protocol to amplify immunofluorescent signals (left panel). In addition, cells in buffer containing sodium azide were pre-incubated on ice with recombinant mouse IL-10 (0.25 µg, Cat. No. 554585) prior to staining with Purified Rat Anti-Mouse CD210 to determine the blocking effect of bound IL-10 protein on staining with 1B1.3a (right panel). Staining with the purified 1B1.3a antibody (filled histograms) is compared to the staining obtained with the secondary antibody alone (open histograms). Histograms in both panels are gated on the lymphocyte population as defined by light scattering characteristics and were also gated on the cell subset stained with Purified Rat Anti-Mouse CD210 (Cat. No. 553088).

Flow cytometric analysis of CD210 expression on mouse splenocytes. Mouse B220+ splenocytes were stained with Purified Rat Anti-Mouse CD210 (Cat. No. 559912) followed by Biotin Goat Anti-Rat Ig (Cat. No. 554014) and Streptavidin-PE (Cat. No. 554061) in a three-layer staining protocol to amplify immunofluorescent signals (left panel). In addition, cells in buffer containing sodium azide were pre-incubated on ice with recombinant mouse IL-10 (0.25 µg, Cat. No. 554585) prior to staining with Purified Rat Anti-Mouse CD210 to determine the blocking effect of bound IL-10 protein on staining with 1B1.3a (right panel). Staining with the purified 1B1.3a antibody (filled histograms) is compared to the staining obtained with the secondary antibody alone (open histograms). Histograms in both panels are gated on the lymphocyte population as defined by light scattering characteristics and were also gated on the cell subset stained with Purified Rat Anti-Mouse CD210 (Cat. No. 553088).

Product Details
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BD Pharmingen™
IL-10 Receptor
Mouse (QC Testing)
Rat IgG1, κ
Purified recombinant ligand-binding domain of mIL-10R
Flow cytometry (Routinely Tested)
0.5 mg/ml
AB_397367
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Recommended Assay Procedures

Immunofluorescent staining and flow cytometric analysis: Purified Rat Anti-Mouse CD210 (Cat. No. 559912) can be used for immmunofluorescent staining (≤ 1 µg antibody/10e6 cells) and flow cytometric analysis of the levels of membrane IL-10R expressed by mouse cell lines or mouse lymphoid cells. It is recommended that the purified format of this antibody be used in conjunction with Biotin Goat Anti-Rat Ig (Cat. No. 554014) and streptavidin-phycoerythrin (PE) (Cat. No. 554061) in a three-layer staining procedure to amplify immunofluorescent signals.

Note: Since 1B1.3a is a neutralizing antibody, it competes with IL-10 for binding to its receptor. Therefore, the use of the 1B1.3a antibody for immunofluorescent staining and flow cytometric analysis in systems where the natural ligand of the receptor is present may give an underestimation of IL-10R expression. Based on our historical testing results, the presence of recombinant mouse IL-10 protein at levels above 250 ng/ml is sufficient to partially inhibit the binding of the 1B1.3a antibody (at 0.12 µg/10e6 cells).

In vitro neutralization: The NA/LE format of this antibody (Cat. No. 550012) is useful for bioassay.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  4. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
  5. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
559912 Rev. 2
Antibody Details
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1B1.3a

The 1B1.3a antibody reacts with the extracellular region of mouse CD210 which is also known as the mouse IL-10 receptor (mIL-10R); it does not recognize the human IL-10R. The IL-10R is expressed by a variety of mouse cell types and cell lines including thymocytes, T cells, B cells, and monocytes. 1B1.3a is a neutralizing antibody and reportedly blocks the binding of human IL-10, which cross-reacts with the mouse IL-10R. The immunogen used for the generation of the 1B1.3a hybridoma was purified recombinant ligand-binding domain of mIL-10R.

559912 Rev. 2
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
559912 Rev.2
Citations & References
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Development References (2)

  1. O'Farrell AM, Liu Y, Moore KW, Mui AL. IL-10 inhibits macrophage activation and proliferation by distinct signaling mechanisms: evidence for Stat3-dependent and -independent pathways. EMBO J. 1998; 17(4):1006-1018. (Immunogen: Neutralization). View Reference
  2. Zola H. Detection of cytokine receptors by flow cytometry. In: Coligan JE, Kruisbeek AM, Margulies DH, Shevach EM, Strober W, ed. Current Protocols in Immunology. New York: Green Publishing Associates and Wiley-Interscience; 1995:6.21.1-6.21.18.
559912 Rev. 2

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.