Western Blotting (WB), ELISA, Immunoprecipitation (IP)
Sequence
DYKDDDDK
Characteristics
Concentration Definition: by UV absorbance at 280 nm
Immunogen
This antibody was purified from whole rabbit serum prepared by repeated immunizations with the Enterokinase (ECS) peptide DYKDDDDK (Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys) conjugated to KLH using maleimide. Residues of glycine and cysteine were added to the carboxy terminal end to facilitate coupling. This antibody reacts with DYKDDDDK conjugated proteins.
This antibody is optimally suited for monitoring the expression of DYKDDDDK tagged fusion proteins. As such, this antibody can be used to identify fusion proteins containing the DYKDDDDK epitope. The antibody recognizes the epitope tag fused to the amino- and carboxy- termini of targeted proteins. This antibody has been tested by ELISA and western blotting against both the immunizing peptide and DYKDDDDK containing recombinant proteins. Although not tested, this antibody is likely functional for immunoprecipitation and immunocytochemistry, and other immunodetection techniques. The epitope tag peptide sequence was first derived from the 11-amino-acid leader peptide of the gene-10 product from bacteriophage T7. Now the most commonly used hydrophilic octapeptide is DYKDDDDK. polyclonal antibody to detect DYKDDDDK conjugated proteins binds DYKDDDDK containing fusion proteins with greater affinity than the widely used monoclonal M1, M2 and M5 clones, and shows greater sensitivity in most assays. Affinity purification of the polyclonal antibody results in very low background levels in assays and low cross-reactivity with other cellular proteins.
Restrictions
For Research Use only
Format
Lyophilized
Reconstitution
Restore with deionized water (or equivalent)
Concentration
1.0 mg/mL
Buffer
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Preservative
Gentamicin sulfate
Handling Advice
Do NOT add Sodium Azide!
Storage
4 °C
Benjamin, van der Meer, Im, Plosa, Zaynagetdinov, Burman, Havrilla, Gleaves, Polosukhin, Deutsch, Yanagisawa, Davidson, Prince, Young, Blackwell: "Epithelial-Derived Inflammation Disrupts Elastin Assembly and Alters Saccular Stage Lung Development." in: The American journal of pathology, Vol. 186, Issue 7, pp. 1786-1800, (2017) (PubMed).
Epitope tags are short peptide sequences that are easily recognized by tag-specific antibodies. Due to their small size, epitope tags do not affect the tagged protein’s biochemical properties. Most often sequences encoding the epitope tag are included with target DNA at the time of cloning to produce fusion proteins containing the epitope tag sequence. This allows anti-epitope tag antibodies to serve as universal detection reagents for any tag containing protein produced by recombinant means. This means that anti-epitope tag antibodies are a useful alternative to generating specific antibodies to identify, immunoprecipitate or immunoaffinity purify a recombinant protein. The anti-epitope tag antibody is usually functional in a variety of antibody-dependent experimental procedures. Expression vectors producing epitope tag fusion proteins are available for a variety of host expression systems including bacteria, yeast, insect and mammalian cells. Supplier produces anti-epitope tag antibodies against many common epitope tags including Myc, GST, GFP, 6X His, MBP, DYKDDDDK and HA. Supplier also produces antibodies to other tags including FITC, Rhodamine (TRITC), DNP and biotin. Synonyms: HRP