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- 文献和实验
- 技术资料
- 供应商:
BD Pharmingen
- 靶点:
CD28
- 级别:
RUO
- 克隆性:
单克隆
- 抗原来源:
Human
- 抗体英文名:
CD28 Antibody
- 抗体名:
CD28
- 标记物:
BV421
- 宿主:
Mouse
- 适应物种:
Human
- 免疫原:
Human CD28 Transfected Cell Line
- 亚型:
Mouse C3H x BALB/c IgG1, κ
- 形态:
液体
- 应用范围:
Flow Cytometry (FCM)
- 保存条件:
2-8℃避光保存
- 规格:
100 Tests
产品概述
Hu CD28 BV421 CD28.2 100Tst 为 BD Pharmingen 品牌流式抗体,货号 562613,靶向 CD28,荧光标记 BV421,克隆 CD28.2 (RUO),规格 100 Tests。适用于 Flow cytometry (Routinely Tested) 检测,仅供科研使用(RUO)。
靶点与抗体说明
The CD28.2 monoclonal antibody specifically binds to CD28, a 44 kDa homodimeric transmembrane glycoprotein present on most mature T cells, thymocytes and plasma cells. CD28 is a costimulatory receptor that binds CD80 and CD86 as ligands and plays a very important role in T cell-B cell interactions. It has been suggested that CD28 initiates and regulates a separate and distinct signal transduction pathway from those stimulated by the TCR complex. Additionally, it has been reported that CD28 antibody clones vary in their ability to stimulate T cells to produce IL-2 and increase intracellular Ca2+ concentration. This finding suggests the existence of functionally distinct subregions on the CD28 molecule. CD28.2 has been demonstrated to bind to the same molecule as clone L293, another CD28 mAb, and has been reported to induce Ca2+ influx in Jurkat T cells. The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.
核心参数
| 货号 | 562613 |
| 品牌 | BD Pharmingen |
| 克隆号 | 克隆 CD28.2 (RUO) |
| 宿主 | Mouse |
| 同种型 | Mouse C3H x BALB/c IgG1, κ |
| 荧光标记 | BV421 |
| 抗原 / 靶点 | CD28 |
| 反应种属 | Human |
| 验证应用 | Flow cytometry (Routinely Tested) |
| 推荐用量 | 5 µl |
| 免疫原 | Human CD28 Transfected Cell Line |
| 规格 | 100 Tests |
| 存储缓冲液 | Aqueous buffered solution containing BSA and ≤0.09% sodium azide. |
荧光染料信息
BV421 The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
保存与运输
建议 2-8℃ 避光保存,请勿冷冻,避免反复冻融。 存储缓冲液:Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
参考文献
June CH, Bluestone JA, Nadler LM, Thompson CB. The B7 and CD28 receptor families. Immunol Today. 1994; 15(7):321-331. (Biology). Kuiper H, Brouwer M, Vermeire S, van Lier R. Analysis of the Workshop CD28 Panel mAb: distinct signalling pathways coupled to CD28. In: Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995:373-374. Nunes J, Klasen S, Franco MD, et al. Signalling through CD28 T-cell activation pathway involves an inositol phospholipid-specific phospholipase C activity. Biochem J. 1993; 293(3):835-842. (Clone-specific: Calcium Flux, (Co)-stimulation, Functional assay). Nunes J, Klasen S, Ragueneau M, et al. CD28 mAbs with distinct binding properties differ in their ability to induce T cell activation: analysis of early and late activation events. Int Immunol. 1993; 5(3):311-315. (Immunogen: Calcium Flux, (Co)-stimulation, Flow cytometry, Functional assay, IC/FCM Block, Immunoprecipitation, Stimulation). Olive D, Cerdan C, Costello R, et al. CD28 and CTLA-4 cluster report. In: Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995:360-370. Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995.
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文献和实验Use a 1L bag for CD3 T cell separation. Add 100 ml of air to the bag. 3) Add 5 x 108 CD3 T cells to the 1L bag in 50-60 ml Buffer 1. 4) Add 4.0 ml (corresponding to 1.6 x 109 Dynabeads®) of washed Dynabeads® CD3⁄CD28
Dynabeads® Mouse T-Activator CD3/CD28 – for physiological activation of mouse T cells
volume of Dynabeads taken from the vial (step 2). 2. Activation of Mouse T Cells 1) Start with 8 × 104 purified T cells in 100-200 µl medium in a 96-well tissue culture plate. 2) Add 2 µl Dynabeads Mouse T-Activator CD3/CD28
上清。检测细胞内的成份时,用 PBS ( PH7.2-7.4 )稀释细胞悬液,细胞浓度达到 100 万 /ml 左右。通过反复冻融,以使细胞破坏并放出细胞内成份。离心 20 分钟左右( 2000-3000 转 / 分)。仔细收集上清。保存过程中如有沉淀形成,应再次离心。 5. 组织标本:切割标本后,称取重量。加入一定量的 PBS , PH7.4 。用液氮迅速冷冻保存备用。标本融化后仍然保持 2-8℃ 的温度。加入一定量的 PBS ( PH7.4 ),用手工或匀浆器将标本匀浆充分。离心
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