PBS, 25% Glycerol, 1% BSA, 0.3% Proclin 300
12 months from date of receipt / reconstitution, -20 °C as supplied
| 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.
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.
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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