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- 详细信息
- 文献和实验
- 技术资料
- 免疫原:
Recombinant human HLA-F protein
- 亚型:
IgG1
- 保存条件:
Store at -80°C.
- 克隆性:
Monoclonal
- 标记物:
Unconjugated
- 适应物种:
human
- 宿主:
Mouse
- 应用范围:
WB, IHC, IF/ICC, Indirect ELISA
- 靶点:
HLA-F
- 抗体英文名:
HLA-F Monoclonal antibody, PBS Only
- 抗体名:
HLA-F Monoclonal antibody, PBS Only
- 规格:
100ug
产品信息
66819-1-PBS targets HLA-F in WB, IHC, IF/ICC, Indirect ELISA applications and shows reactivity with human samples.
| 经测试应用 | WB, IHC, IF/ICC, Indirect ELISA |
| 经测试反应性 | human |
| 免疫原 |
CatNo: Ag6475 Product name: Recombinant human HLA-F protein Source: e coli.-derived, PET28a Tag: 6*His Domain: 21-298 aa of BC062991 Sequence: AGSHSLRYFSTAVSRPGRGEPRYIAVEYVDDTQFLRFDSDAAIPRMEPREPWVEQEGPQYWEWTTGYAKANAQTDRVALRNLLRRYNQSEAGSHTLQGMNGCDMGPDGRLLRGYHQHAYDGKDYISLNEDLRSWTAADTVAQITQRFYEAEEYAEEFRTYLEGECLELLRRYLENGKETLQRADPPKAHVAHHPISDHEATLRCWALGFYPAEITLTWQRDGEEQTQDTELVETRPAGDGTFQKWAAVVVPSGEEQRYTCHVQHEGLPQPLILRWEQS |
| 宿主/亚型 | Mouse / IgG1 |
| 抗体类别 | Monoclonal |
| 产品类型 | Antibody |
| 全称 | major histocompatibility complex, class I, F |
| 别名 | 1B12C2, CDA12, HLA 5.4, HLA CDA12, HLA F |
| 计算分子量 | 39 kDa |
| 观测分子量 | 40-45 kDa |
| GenBank蛋白编号 | BC062991 |
| 基因名称 | HLA-F |
| Gene ID (NCBI) | 3134 |
| RRID | AB_2882162 |
| 偶联类型 | Unconjugated |
| 形式 | Liquid |
| 纯化方式 | Protein G purification |
| UNIPROT ID | P30511 |
| 储存缓冲液 | PBS only, pH 7.3. |
| 储存条件 | Store at -80°C. The product is shipped with ice packs. Upon receipt, store it immediately at -80°C |
背景介绍
Human major histocompatibility complex (MHC) antigens, also referred to as human leukocyte antigens (HLA), are encoded by genes located on the short arm of chromosome 6 (6p21.3). There are two classes of HLA antigens: class I and class II. This class I molecules are membrane glycoproteins composed of a heavy (alpha) chain which is encoded by a HLA class I gene, and β2-microglobulin light (beta) chain. The most extensively characterized members of the HLA class I gene family are the genes encoding the major transplantation antigenes, HLA-A, B and C. HLA-F is a non-classical MHC class I molecule. (PMID: 667938; 3375250; 2249951)
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文献和实验1. 杂交瘤阳性克隆的筛选与克隆化 在HAT培养基中生长的杂交瘤细胞,只有少数是分泌预定特异性单克隆抗体的细胞,因此,必须进行筛选和克隆化。通常采用有限稀释法进行杂交瘤细胞的克隆化培养。采用灵敏、快速、特异的免疫学方法,筛选出能产生所需单克隆抗体的阳性杂交瘤细胞,并进行克隆扩增。经过全面鉴定其所分泌单克隆抗体的免疫球蛋白类型、亚类、特异性、亲和力、识别抗原的表位及其分子量后,及时进行冻存。2. 单克隆抗体的大量制备 单克隆抗体的大量制备重要采用动物体内诱生法和体外培养法。
方法1、抗体测定:研究对象取200μl新鲜抗凝全血,每管100μl加入流式细胞仪检测管中,再加入2ml红细胞裂解液反应12min后离心,弃上清液,磷酸盐缓冲液(phosphate buffered saline, PBS)洗细胞两次,其中反应管加入鼠抗人HLA-A*02抗体××μl(达科为公司),4℃ 30min孵育后,PBS再次洗细胞,每管加入PE标记羊抗鼠免疫球蛋白2μl,4℃、30
过夜。(5)用少许PBS将交联物全部洗出,收集洗出液,加1/20V体积新鲜配制的5mg/ml NaBH4溶液,混匀,室温作用30分钟;再加入3/20V NaBH4溶液,混匀,室温作用1小时(或4℃过夜)。(6)将交联物过Sephadex G200或Sepharose 6B(2.6×50cm)层析纯化,分管收集第一峰。(7)酶结合物质量鉴定:克分子比值测定酶量(mg/ml)=OD403×0.4IgG量(mg/ml)=(OD280-OD403×0.3)×0.62克分子比值(E/P)=酶量×4/IgG量
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