DYKDDDDK Epitope Tag Rabbit Polyclonal Antibody

Specifications

Product Data
Applications	ELISA, IHC, WB
Recommend Dilution	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 either the amino- or 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. This 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. Recommended Dilutions: ELISA: 1/90,000-1/250,000. Western blot: 1/2,000-1/10,000. Immunohistochemistry:  User-optimized
Host	Rabbit
Clonality	Polyclonal
Immunogen	This antibody was purified from whole rabbit serum prepared by repeated immunizations with the DYKDDDDK epitope tag peptide (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.
Specificity	This affinity purified antibody is directed against the DYKDDDDK motif and is useful in determining its presence in various assays. This polyclonal anti-DYKDDDDK tag antibody detects over-expressed proteins containing the DYKDDDDK epitope tag. To date this antibody has reacted with all amino-terminal DYKDDDDK tagged proteins so far tested. In western blotting of bacterial extracts the antibody does not cross-react with endogenous proteins.
Formulation	0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2 State: Aff - Purified State: Liquid (sterile filtered) purified Ig fraction Preservative: 0.01% Sodium Azide
Concentration	lot specific
Purification	Affinity purification of this polyclonal antibody results in very low background levels in assays and low cross-reactivity with other cellular proteins.
Conjugation	Unconjugated
Storage Condition	Store the antibody undiluted at 2-8°C for one month or (in aliquots) at -20°C or below for longer. Avoid repeated freezing and thawing. Dilute only prior to immediate use.
Background	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. 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.
Synonyms	FLAG Epitope Tag, ECS Epitope Tag, FLAG-tag, ECS-tag, D-tag
Reference Data

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