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Flow cytometric profile of CD210a expression on human peripheral blood lymphocytes. Human whole blood was stained with either Purified Rat IgG2a, κ Isotype Control (Cat. No. 555841; dashed line histogram) or Purified Rat Anti-Human CD210a (Cat. No. 556012; solid line histogram). Three-step staining was carried out with Biotin Goat Anti-Rat Ig (Cat. No. 554014) and PE Streptavidin (Cat. No. 554061). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed on a BD FACSCan™ System.
BD Pharmingen™ Purified Rat Anti-Human CD210a
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
For flow cytometric applications, a three step labeling procedure is recommended for amplifying signal. Suggested protocol for 3-step staining using Lysed Whole Blood method:
1. Incubate 100µl whole blood with primary (unconjugated) antibody for 20-30 minutes in the dark at room temperature.
2. Add 2 mls of 1X Lysing Buffer (Cat. No. 555899/349202) and incubate for 10-15 minutes in the dark. Centrifuge and aspirate.
3. Wash once with PBS/0.1% sodium azide/1% heat-inactivated fetal bovine serum (PBS-FBS). Centrifuge and aspirate.
4. Add biotinylated goat anti-rat Ig's and incubate for 20-30 minutes in the dark at room temperature.
5. Wash once with PBS-FBS. Centrifuge and aspirate.
6. Add SAV-PE (Cat. No. 554061) and incubate for 20-30 minutes in the dark at room temperature.
7. Wash once with PBS-FBS. Centrifuge and aspirate. Resuspend in 0.5 ml of PBS-FBS and analyze by flow cytometry.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The 3F9 monoclonal antibody specifically binds to CD210a, which is also known as, Interleukin-10 Receptor subunit alpha (IL-10R subunit alpha/IL-10Rα/IL10RA), or Interleukin-10 receptor subunit 1 (IL-10R1). CD210a is a 90-110 kDa type I transmembrane glycoprotein that belongs to the type II cytokine receptor family. CD210a combines with IL-10 Receptor subunit beta (IL-10Rβ/IL10RB/CDw210b) to form the IL-10 Receptor complex (IL-10Rα/IL-10Rβ) that can bind IL-10 and transduce signals intracellularly through the JAK/STAT pathway. IL-10 can suppress antigen presentation and the expression of proinflammatory type-1 immune responses while promoting type-2 immune responses. CD210a is expressed on T cells, B cells, NK cells, monocyte, macrophages and dendritic cells. Clone 3F9 is specific for human CD210a and its binding to the receptor can be blocked by recombinant human IL-10 (rhIL-10) protein.
Development References (4)
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Callard R, Gearing A. Callard R, Gearing A. The Cytokine Facts Book. San Diego: Academic Press; 1994.
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Carson WE, Lindemann MJ, Baiocchi R, et al. The functional characterization of interleukin-10 receptor expression on human natural killer cells. Blood. 1995; 85(12):3577-3585. (Biology). View Reference
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Liu Y, Wei SH, Ho AS, de Waal Malefyt R, Moore KW. Expression cloning and characterization of a human IL-10 receptor. J Immunol. 1995; 152(4):1821-1829. (Biology). View Reference
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Liu Y, de Waal Malefyt R, Briere F, et al. The EBV IL-10 homologue is a selective agonist with impaired binding to the IL-10 receptor.. J Immunol. 1997; 158(2):604-13. (Biology). View Reference
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.