This product is an IgG fraction antibody purified from ascites by Protein A chromatography followed by extensive dialysis against the buffer stated above. The purified antibody is directed against the DYKDDDDK motif and is useful in determining its presence in various assays where the epitope tag is present at either the amino or carboxy terminus of recombinant proteins. This monoclonal anti-DYKDDDDK™ tag antibody detects over-expressed proteins containing the DYKDDDDK epitope tag. In western blotting of bacterial extracts, the antibody does not cross-react with endogenous proteins.
Characteristics
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.
Sterility
Sterile filtered
Immunogen
This antibody was produced in mice by repeated immunizations with a synthetic peptide corresponding to the DYKDDDDK epitope tag peptide DYKDDDDK (Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys) conjugated to KLH using maleimide. Residues of glycine and cysteine were added to the termini to facilitate coupling. Immunogen Type: Peptide
WB, ELISA, IP
Host: Mouse
Monoclonal
5A8E5
unconjugated
Application Notes
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. The epitope tag peptide sequence was first derived from the 11-amino-acid leader peptide of the gene-10 product from bacteriophage T7. DYKDDDDK is the most commonly used hydrophilic octapeptide tag.
Restrictions
For Research Use only
Format
Liquid
Concentration
1.0 mg/mL
Buffer
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Preservative
Sodium azide
Precaution of Use
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage
4 °C/-20 °C
Storage Comment
Store vial at 4 °C prior to restoration. For extended storage aliquot contents and freeze at -20 °C or below. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4 °C as an undiluted liquid. Dilute only prior to immediate use. Expiration date is one (1) year from date of opening.
Expiry Date
12 months
Huang, Chuang, Hao, Talbot, Sen, Trink, Sidransky, Ratovitski: "Phospho-?Np63? is a key regulator of the cisplatin-induced microRNAome in cancer cells." in: Cell death and differentiation, Vol. 18, Issue 7, pp. 1220-30, (2011) (PubMed).
Chubet, Brizzard: "Vectors for expression and secretion of FLAG epitope-tagged proteins in mammalian cells." in: BioTechniques, Vol. 20, Issue 1, pp. 136-41, (1996) (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: DYKDDDDK tag, Enterokinase Cleavage Site (ECS), DYKDDDDK, Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys