FLAG Tag Knockin Mice
Since 1998, ingenious targeting laboratory has generated FLAG tag knockin models enabling detection and purification of endogenous proteins without requiring gene specific antibodies.
The FLAG epitope (DYKDDDDK) provides reliable detection across multiple applications with widely available, well characterized antibodies. FLAG tag knockin at endogenous loci preserves physiological expression levels while providing standardized reagents for protein detection, immunoprecipitation, and affinity purification.
Frequently asked questions
FLAG tag is inserted at the N-terminus (after start codon) or C-terminus (before stop codon) of the target gene, creating a fusion protein. The FLAG tag sequence (DYKDDDDK) is inserted in-frame. N-terminal tagging is common for immunoprecipitation; C-terminal tagging preserves signal sequences.
FLAG tag provides excellent antibody availability with high-affinity anti-FLAG antibodies suitable for immunoprecipitation, Western blot, immunohistochemistry, and flow cytometry. FLAG tag is small (8 amino acids), minimizing potential effects on protein function. It offers high specificity with minimal cross-reactivity.
Yes. Conditional FLAG tag knockins can use LoxP-flanked stop cassettes that prevent tag expression until Cre-mediated recombination removes the stop. This enables tissue specific or temporal control of tagged protein expression when combined with appropriate Cre driver lines.
Pre-germline characterization includes Southern blot analysis to confirm correct targeting and sequence verification to ensure FLAG tag sequence integrity. Post-germline validation includes Western blot (anti-FLAG antibody), immunoprecipitation, or immunohistochemistry to confirm tag expression and proper protein localization.
