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宿主来源
Rabbit抗原名称
FLAG Tag免疫原
Synthetic Peptide克隆号
S-589-8抗体类型
Recombinant mAb抗体同种型
IgG反应种属 ?
Species Independent阳性样本
H3-Flag-V5 transfected 293T cells纯化方式
Protein A浓度
2 mg/ml标记
FITC性状
Liquid缓冲体系
PBS, 1% BSA, 0.3% Proclin 300储存条件
12 months from date of receipt / reconstitution, 2 to 8 °C as supplied.
应用
ICFCM ,ICC ?稀释度
应用 稀释度 推荐种属 ICC 1:500 Species Independent ICFCM 1:800 Species Independent
FLAG-tag, or FLAG octapeptide, or FLAG epitope, is a peptide protein tag that can be added to a protein using recombinant DNA technology, having the sequence DYKDDDDK (where D=aspartic acid, Y=tyrosine, and K=lysine). It is one of the most specific tags and it is an artificial antigen to which specific, high affinity monoclonal antibodies have been developed and hence can be used for protein purification by affinity chromatography and also can be used for locating proteins within living cells. FLAG-tag has been used to separate recombinant, overexpressed protein from wild-type protein expressed by the host organism. FLAG-tag can also be used in the isolation of protein complexes with multiple subunits.
流式分析
Flag Tag transfected 293T (Human embryonic kidney epithelial cell, Right panel) or 293T (Left panel) was fixed with 4% PFA and permeabilized 90% methanol ，then cells were stained with either FITC Rabbit IgG Isotype Control (Black line histogram) or SDT FLAG Tag Recombinant Rabbit mAb (FITC Conjugate) (S-589-8) (Red line histogram) at 1/800 dilution (0.25 μg), cells without incubation with primary antibody and secondary antibody (Blue line histogram) was used as unlabelled control. Flow cytometry and data analysis were performed using BD FACSymphony™ A1 and FlowJo™ software.
免疫细胞化学
ICC shows positive staining in Histone H3-Flag-V5 transfected 293T cells (top panel) and negative staining in vector-transfected 293T cells (below panel). Anti- FLAG Tag (FITC Conjugate) antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. The cells were fixed with 100% ice-cold methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
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