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PE Rat Anti-Human and Viral IL-10
PE Rat Anti-Human and Viral IL-10

Expression of IL-10 by stimulated CD14+ human monocytes. Human PBMC were stimulated for 24 hr with LPS (1.0 µg/ml) in the presence of BD GolgiStop™ (Cat No. 554724). The PBMC were stained with FITC-mouse anti-human CD14 antibody (FITC-M5E2, Cat. No. 555397) and 0.25 µg of PE-rat anti-human IL-10 antibody (PE-JES3-9D7, Cat. No. 554498) by using BD Biosciences Pharmingen's staining protocol (see left panel). The data reflect gating on monocytes, based on forward and side scatter. The binding of PE-JES3-9D7 was blocked by the preincubation of the conjugate with an excess of rhIL-10 (1 mg; Cat. No. 554611) (see right panel). The quadrant markers for the bivariate dot plots were set based on the unstained cell control.

Expression of IL-10 by stimulated CD14+ human monocytes. Human PBMC were stimulated for 24 hr with LPS (1.0 µg/ml) in the presence of BD GolgiStop™ (Cat No. 554724). The PBMC were stained with FITC-mouse anti-human CD14 antibody (FITC-M5E2, Cat. No. 555397) and 0.25 µg of PE-rat anti-human IL-10 antibody (PE-JES3-9D7, Cat. No. 554498) by using BD Biosciences Pharmingen's staining protocol (see left panel). The data reflect gating on monocytes, based on forward and side scatter. The binding of PE-JES3-9D7 was blocked by the preincubation of the conjugate with an excess of rhIL-10 (1 mg; Cat. No. 554611) (see right panel). The quadrant markers for the bivariate dot plots were set based on the unstained cell control.

Product Details
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BD Pharmingen™
Human (QC Testing), Viral (Tested in Development)
Rat IgG1
Recombinant Human IL-10
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
AB_395434
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

Recommended Assay Procedures

For optimal immunofluorescent staining with flow cytometric analysis, this anti-cytokine antibody should be pretitrated (≤ 0.50 µg mAb/million cells). An appropriate isotype control for this antibody for intracellular staining is PE-conjugated rat IgG1, clone R3-34 (Cat. No. 554685). For specific methodology, please visit the protocols section or chapter on intracellular staining in the Immune Function Handbook, both of which are posted on our web site, www.bdbiosciences.com.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  4. 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.
554498 Rev. 3
Antibody Details
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JES3-9D7

The JES3-9D7 monoclonal antibody specifically reacts with human IL-10 (Interleukin-10) and viral IL-10. The immunogen used to generate the JES3-9D7 hybridoma was recombinant human IL-10 expressed by COS cells. IL-10 is also known as CSIF (Cytokine synthesis inhibitory factor) and (TGIF) T-cell growth inhibitory factor. IL-10 is expressed by various cell types including activated monocytes, macrophages, dendritic cells, mast cells, granulocytes, and lymphocytes. IL-10 is a pleiotropic cytokine that can downregulate proinflammatory immune responses, such as Th1-like responses, while promoting other responses including B cell proliferation and antibody production.

554498 Rev. 3
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
554498 Rev.3
Citations & References
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Development References (6)

  1. Abrams J. Immunoenzymetric assay of mouse and human cytokines using NIP-labeled anti-cytokine antibodies. Curr Protoc Immunol. 2001; 1:6.20-6.21. (Clone-specific: ELISA). View Reference
  2. Abrams JS, Roncarolo MG, Yssel H, Andersson U, Gleich GJ, Silver JE. Strategies of anti-cytokine monoclonal antibody development: immunoassay of IL-10 and IL-5 in clinical samples. Immunol Rev. 1992; 127:5-24. (Clone-specific: ELISA). View Reference
  3. Burdin N, Peronne C, Banchereau J, Rousset F. Epstein-Barr virus transformation induces B lymphocytes to produce human interleukin 10. J Exp Med. 1993; 177(2):295-304. (Clone-specific: ELISA). View Reference
  4. Gotlieb WH, Abrams JS, Watson JM, Velu TJ, Berek JS, Martinez-Maza O. Presence of interleukin 10 (IL-10) in the ascites of patients with ovarian and other intra-abdominal cancers. Cytokine. 1992; 4(5):385-390. (Clone-specific: ELISA). View Reference
  5. Prussin C, Metcalfe DD. Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies. J Immunol Methods. 1995; 188(1):117-128. (Methodology: Flow cytometry). View Reference
  6. Yssel H, De Waal Malefyt R, Roncarolo MG, et al. IL-10 is produced by subsets of human CD4+ T cell clones and peripheral blood T cells. J Immunol. 1992; 149(7):2378-2384. (Clone-specific: ELISA). View Reference
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554498 Rev. 3

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