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BD Pharmingen- PE Mouse Anti-Rat IL-10_BD Pharmingen

艾美捷科技 • 2025年08月22日 15:26:16 • 抗体 • 阅读 54 • 标签
产品信息
荧光素标记
PE
抗原名称
IL-10
宿主
Mouse IgG2b, κ
免疫原
Recombinant Rat IL-10
简单描述
The A5-4 monoclonal antibody specifically binds to rat interleukin-10 (IL-10). The immunogen used to generate the A5-4 hybridoma was recombinant rat IL-10 protein.
商品描述
A5-4 The A5-4 monoclonal antibody specifically binds to rat interleukin-10 (IL-10). The immunogen used to generate the A5-4 hybridoma was recombinant rat IL-10 protein.
同种型
Mouse IgG2b, κ
克隆号
克隆 A5-4 (RUO)
浓度
0.2 mg/ml
产品详情
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.
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
应用
实验应用
Intracellular block/flow cytometry (Routinely Tested)
反应种属
Rat (QC Testing)
目标/特异性
IL-10
背景
别名
Il10; Interleukin-10; CSIF; Cytokine synthesis inhibitory factor
制备和贮存
存储溶液
Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
保存方式
Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
文献
文献
研发参考(2) 1. Feng L, Tang WW, Chang JC, Wilson CB. Molecular cloning of rat cytokine synthesis inhibitory factor (IL-10) cDNA and expression in spleen and macrophages. Biochem Biophys Res Commun. 1993; 192(2):452-458. (Immunogen). 2. 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: IC/FCM Block).

参考图片

Expression of IL-10 by stimulated CD4+ rat splenocytes. Purified splenic CD4+ cells from 6 month old Lewis rats were stimulated with plate-bound anti-CD3 (25 µg/ml final concentration; G4.18, Cat. No. 554829) and soluble anti-rat CD28 (2 µg/ml final concentration; clone JJ319, Cat. No. 554993) for 2 days in culture together with rat IL- 2 (10 ng/ml final concentration; Cat. No. 555106) and rat IL-4 (10 ng/ml final concentration; Cat. No 555107), followed by a 3 day incubation with only rat IL-2 and rat IL-4. This was followed by a 6 hour stimulation with PMA (1 ng/ml final concentration; Sigma, Cat. #P-8139) and Ionomycin (250 ng/ml final concentration; Sigma, Cat. #I-0634) in the presence of BD GolgiStop ™ (2 µM final concentration Cat. No. 554724). The cells were then fixed, permeabilized, and subsequently stained with 0.06 µg of PE Mouse anti rat IL-10 antibody (PE-A5-4, Cat. No. 555088; Left panel) by using Pharmingen's staining protocol. To demonstrate specificity of staining, the binding of the PE-A5-4 antibody was blocked by preincubation of the PE- conjugated antibody with recombinant rat IL-10 (0.25 µg, Cat. No. 555113; Second panel), and by preincubation of the fixed/permeabilized cells with the unlabeled A5-4 antibody (10 µg; Right panel) prior to staining. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control, and verified with the recombinant cytokine blocking (Center panel) and unlabeled antibody blocking (Right panel) specificity controls.

Expression of IL-10 by stimulated CD4+ rat splenocytes. Purified splenic CD4+ cells from 6 month old Lewis rats were stimulated with plate-bound anti-CD3 (25 µg/ml final concentration; G4.18, Cat. No. 554829) and soluble anti-rat CD28 (2 µg/ml final concentration; clone JJ319, Cat. No. 554993) for 2 days in culture together with rat IL- 2 (10 ng/ml final concentration; Cat. No. 555106) and rat IL-4 (10 ng/ml final concentration; Cat. No 555107), followed by a 3 day incubation with only rat IL-2 and rat IL-4. This was followed by a 6 hour stimulation with PMA (1 ng/ml final concentration; Sigma, Cat. #P-8139) and Ionomycin (250 ng/ml final concentration; Sigma, Cat. #I-0634) in the presence of BD GolgiStop ™ (2 µM final concentration Cat. No. 554724). The cells were then fixed, permeabilized, and subsequently stained with 0.06 µg of PE Mouse anti rat IL-10 antibody (PE-A5-4, Cat. No. 555088; Left panel) by using Pharmingen's staining protocol. To demonstrate specificity of staining, the binding of the PE-A5-4 antibody was blocked by preincubation of the PE- conjugated antibody with recombinant rat IL-10 (0.25 µg, Cat. No. 555113; Second panel), and by preincubation of the fixed/permeabilized cells with the unlabeled A5-4 antibody (10 µg; Right panel) prior to staining. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control, and verified with the recombinant cytokine blocking (Center panel) and unlabeled antibody blocking (Right panel) specificity controls.

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