Epitope tagging is a powerful and versatile strategy for detecting and purifying proteins expressed by cloned genes. To utilize this feature, protein expression vectors are typically engineered with a nucleotide sequence that encodes the peptide epitope tag. The gene of interest is cloned in-frame relative to the tag and, upon expression, the protein of interest is synthesized as a fusion protein with the peptide tag. Fusion protein detection and/or purification is mediated by highly specific antibodies to the engineered peptide, thus eliminating the need for antibodies to proteins from each newly cloned gene.
Applications
Suitable for use in Western Blot. Other applications have not been tested.
Recommended Dilution
Western Blot: 1:10,000-1:30,000 when detected with an anti-chicken horseradish peroxidase-conjugated secondary by ECL or by a colorimetric assay using DAB. Optimal dilutions to be determined by the researcher.
Recommended Secondary Antibodies
I1906: IgY, Chicken Pab Rb x Ch I1906-10: IgY, Chicken (HRP) Rb x Ch I1906-12: IgY, Chicken (FITC) Rb x Ch I1906-14: IgY, Chicken (AP) Rb x Ch
Storage and Stability
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Immunogen
Recombinant red fluorescent protein (RFP), highly purified by affinity chromatography on Nickel column.
Form
Supplied as a liquid in 10mM Tris, pH 7.5, 100mM sodium chloride, 0.02% sodium azide.
Purity
Purified by affinity chromatography.
Specificity
Recognizes red fluorescent protein (RFP). Shows no reactivity to GFP.