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- 文献和实验
- 技术资料
- 供应商:
BD Pharmingen
- 靶点:
CD45
- 级别:
RUO
- 克隆性:
单克隆
- 抗原来源:
Human
- 抗体英文名:
CD45 Antibody
- 抗体名:
CD45
- 标记物:
V500
- 宿主:
Mouse
- 适应物种:
Human
- 免疫原:
Human Peripheral Blood Leucocytes
- 亚型:
Mouse 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 CD45 Horizon V500 HI30 100Tst 为 BD Pharmingen 品牌流式抗体,货号 560777,靶向 CD45,荧光标记 V500,克隆 HI30 (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 HI30 monoclonal antibody specifically binds to the 180, 190, 205, 220 kDa protein isoforms of CD45. CD45 is encoded by the PTPRC ( Protein tyrosine phosphatase receptor type C) gene. CD45, also known as the leukocyte common antigen (LCA), is a member of the protein tyrosine phosphatase (PTP) family. It is present on all human leukocytes including lymphocytes, monocytes, granulocytes, eosinophils, and thymocytes. CD45 is absent from circulating erythrocytes, platelets, or mature erythroid cells of bone marrow and non-hemopoietic tissues. The antibody is conjugated to BD Horizon™ V500, which has been developed for use in multicolor flow cytometry experiments and is available exclusively from BD Biosciences. It is excited by the Violet laser with an Ex max of 415 nm and Em Max at 500 nm. BD Horizon V500 conjugates emit at a similar wavelength to Amcyan yet exhibit reduced spillover into the FITC channel. For more information on BD Horizon V500, visit bdbiosciences.com/colors. When compensating dyes in this spectral range (such as Horizon™ V500 and AmCyan), the most accurate compensation can be obtained using single stained cellular controls. Due to spectral differences between cells and beads in this channel, using BD CompBeads can result in spillover errors for V500 and AmCya
核心参数
| 货号 | 560777 |
| 品牌 | BD Pharmingen |
| 克隆号 | 克隆 HI30 (RUO) |
| 宿主 | Mouse |
| 同种型 | Mouse IgG1, κ |
| 荧光标记 | V500 |
| 抗原 / 靶点 | CD45 |
| 反应种属 | Human |
| 验证应用 | Flow cytometry (Routinely Tested) |
| 推荐用量 | 5 µl |
| 免疫原 | Human Peripheral Blood Leucocytes |
| 规格 | 100 Tests |
| 存储缓冲液 | Aqueous buffered solution containing protein stabilizer, glycerol and ≤0.09% sodium azide. |
荧光染料信息
V500 BD Horizon™ V500 Dye is part of the BD Horizon™ Violet family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 415-nm and an emission maximum (Em Max) at 499-nm. Driven by BD innovation, V500 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 500 nm (eg, a 525/50 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye. 格式:V500最多激发源:415 nm最多发射源:499 nm
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 protein stabilizer, glycerol and ≤0.09% sodium azide.
参考文献
Bradstock KF, Janossy G, Pizzolo G, et al. Subpopulations of normal and leukemic human thymocytes: an analysis with the use of monoclonal antibodies. J Natl Cancer Inst. 1980; 65(1):33-42. (Biology). Hermiston ML, Xu Z, Weiss A. CD45: a critical regulator of signaling thresholds in immune cells. Annu Rev Immunol. 2003; 21:107-137. (Biology). Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182. Loken MR, Brosnan JM, Bach BA, Ault KA. Establishing optimal lymphocyte gates for immunophenotyping by flow cytometry. Cytometry. 1990; 11(4):453-459. (Biology). Terry LA, Brown MH, Beverley PC. The monoclonal antibody, UCHL1, recognizes a 180,000 MW component of the human leucocyte-common antigen, CD45. Immunology. 1988; 64(2):331-336. (Biology). Terstappen LW, Levin J. Bone marrow cell differential counts obtained by multidimensional flow cytometry. Blood Cells. 1992; 18(2):311-330. (Biology). Trowbridge IS, Thomas ML. CD45: an emerging role as a protein tyrosine phosphatase required for lymphocyte activation and development. Annu Rev Immunol. 1994; 12:85-116. (Biology). Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.
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文献和实验: 由此步骤开始,流出的溶液包含目标细胞,请使用新的试剂管收集!首先,加入 1 ml 的 biotin 洗脱液,孵育 2 min。其次,加入 10 ml biotin 洗脱液,洗脱目标细胞。d) 再加入 10 ml biotin 洗脱液,洗下剩余的目标细胞。最后两步骤所收集的液体含分选出的目标细胞。(4) 结果分析以下流式图为从 7.5 ml 全血中,选出 CD3+ T 细胞的数据。选后细胞以 CD3-APC(OKT-3) 及 CD45-PE(HI30,白细胞标志) 抗体染色后,进行流式分析,死细胞
- 3) 及 CD45 -PE (HI30,白细胞标志) 抗体染色后,进行流式分析,死细胞以 DAPI 染色先行排除,左右分别为分选前后的细胞群分布。 总结: 几次实验的结果,使用 Fab-TACS® Gravity column 来从全血直接进行 CD3+ T 细胞分选,平均得率为 84.5%,纯度大于 90%,细胞存活率高于 95%,且目标细胞不含标记,贴近天然,使下游实验更精确。
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