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如何选择合适的流式荧光抗体？
选择错误的抗体可能会浪费时间、金钱和宝贵的样本材料，并可能导致发布不正确或不可复制的数据。选择合适的抗体，研究人员应考虑抗体形式、验证应用和抗原靶标的性质等因素，以增加实验成功的可能性。
在选择用于流式细胞术的抗体时，有许多常见的错误。这些错误包括选择识别错误表位的抗体，例如当目的是避免细胞渗透时，选择靶向细胞内表位的产品；或者选择了错误的克隆，或者选择了不合适的荧光素标记或者不合适的荧光配色方案；研究人员在选择用于流式细胞仪的抗体时还应该考虑的另一个因素是使用直标一抗还是间标方法，目前流式实验直标一抗是首选，此外，直标抗体简化了实验步骤，减少了样品制备过程中发生错误的可能性；目前流式应用主要还是选择单克隆抗体，重组抗体也在渐渐进入大家的选择。这是因为多克隆更容易与靶标以外的蛋白质发生交叉反应，并且经常表现出不同的批次间性能。单克隆抗体，尤其是重组抗体，是一致的，可重复的。一般来说，使用重组产生的抗体可以确保真正的克隆性，减少脱靶结合的机会，并提供尽可能高的批次间再现性。
许多研究人员选择使用与FcγR结合的抗体，从而对所有表达这些受体的细胞进行额外染色。例如，同种型如小鼠IgG2a/b和大鼠IgG2b可通过结合巨噬细胞、单核细胞或b细胞而大大增加背景染色。虽然您可以通过在样品制备中添加Fc阻断剂或血清来解决这个问题，但这可能会导致与其他试剂不兼容。例如，在使用最广泛使用的小鼠CD16/32阻断剂进行治疗后，您无法再对大鼠抗体进行染色，因为阻断剂也是大鼠抗体。小片段F（ab）2抗体没有问题。
有这么多抗体可用，选择合适的产品往往是最重要的。凯靖生物10多年的一级和二级抗体生产经验，专注于抗原抗体的研发和生产，公司构建了基于全自动化高通量杂交瘤和单B细胞技术的单克隆抗体对发现平台，为客户提供创优产品和创新服务。是您值得信赖的现成和定制抗体产品合作伙伴。我们的客户包括许领先的体外诊断制造商、制药公司和研究机构，凭借多年客户服务和经验积累，我们提供始终如一的高质量产品。
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| 描述 | CD8, also known as Leu-2 or T8 in human and Lyt2 or Lyt3 in mouse, is a cell surface glycoprotein belonging to the immunoglobulin supergene family. CD8 is expressed on cytotoxic T-lymphocytes (T-cells), most thymocytes, between 35-45% of peripheral blood lymphocytes, and a population of natural killer (NK) cells. The CD8 molecule consists of disulfide-linked alpha (alpha) and beta (beta) chains that present on T-cells as either CD8alphaalpha homodimers or CD8alphabeta heterodimers. Both alpha and beta chains consist of a signaling sequence, an extracellular Ig-like domain, a membrane proximal stalk region, a transmembrane domain, and a cytoplasmic tail. Human CD8alpha is processed as 235 amino acids (aa) in length with a theoretical molecular weight of ~26 kDa, while mouse CD8alpha is 247 aa and has a theoretical molecular weight of 27.5 kDa. Functionally, CD8 acts as an antigen coreceptor on cytotoxic T-cells and interacts with the major histocompatibility complex (MHC) class I molecules on antigen presenting cells (APCs), mediating cell-cell interactions within the immune system. Conversely, CD4 molecules interact with antigens presented on MHC class II molecules and are activated to become helper T-cells (TH). Interestingly, thymocytes can transiently express both CD4 and CD8 during the maturation process. Furthermore, the cytoplasmic tail of CD8 has a Lck (lymphocyte-specific protein tyrosine kinase) binding domain where Lck interacts with CD8, initiating a phosphorylation cascade that activates transcription factors and promotes T-cell activation. More specifically, CD8alphabeta functions as a T-cell co-receptor, while CD8alphaalpha promotes T-cell survival and differentiation. Given its role in the immune system, CD8-deficiency in T-cells is a hallmark of many diseases and pathologies. Specifically, CD8+ T-cell deficiency is prevalent in chronic autoimmune diseases including multiple sclerosis, rheumatoid arthritis, ulcerative colitis, Crohn's disease, type 1 diabetes mellitus, and Graves' disease. Furthermore, cancers or chronic infection can lead to CD8 T-cell exhaustion as the continual antigen presentation and inflammatory signals eventually cause the CD8+ T-cells to lose functionality. However, animal models and clinical studies have suggested that T-cells are capable of being reinvigorated using inhibitory receptor blockade resulting in better disease outcomes and these exhausted T-cells may be a potential therapeutic target. |
| 标记 | percp |
| 克隆号 | KJ3C4 |
| 来源宿主 | mouse |
| 克隆类型 | monoclonal |
| 同种型 | IgG2b |
| 特异性 | human |
| 纯化方式 | Affinity |
| 激发器(nm) | 488 |
| 激发光(ex/nm) | 482 |
| 发射光(em/nm) | 675 |
| 存储溶液 | PBS、0.5%BSA、0.03%PC-300 |
| 存储方式 | Store at 4℃ for one year. |
| 用法用量 | 10 μl/Test, 0.1 mg/ml |
| 使用方法 | The procedure is detailed in the document. |
参考结果：
Typical results from human peripheralblood lymphocytes stained either with#K1011 used at 10 ul/test.
技术支持
datasheet
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