Mouse Monoclonal Antibody [Clone ID: 29E4.G7]

CAT#: TA396761

DYKDDDDK Antibody same epitope as Sigma's Anti-FLAG

Product Images

Twenty-four (24) clones were randomly selected and grown up from glycerol stocks by inoculating 0.5mL 2xYT medium. Expression of recombinant proteins was induced by the addition of IPTG. Proteins were purified by nickel affinity chromatography and eluted in 40 µL. Samples were diluted 10-fold, transferred to nitrocellulose membrane and blotted using Mab-anti-FLAG™ antibody. Personal Communication: A. Morrison and B. Kloss, NYCOMPS, New York, NY.

Specifications

Product Data

• Clone Name: 29E4.G7
• Applications: ELISA, FC, IHC, WB
• Recommended Dilution: WB: 1:2,000-1:10,000
  IHC: 1:1,000-1:5,000
  FC: User Optimized
  ELISA: 1:150,000 - 1:250,000
• Host: Mouse
• Isotype: IgG2a, kappa
• Clonality: Monoclonal
• Immunogen: This antibody was produced in mice by repeated immunizations with a synthetic peptide corresponding to the FLAG™ epitope tag peptide DYKDDDDK (Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys) conjugated to KLH using maleimide.
• Specificity: 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 FLAG™ 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-FLAG™ tag antibody detects over-expressed proteins containing the FLAG™ epitope tag. 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
• Concentration: 1.0 mg/ml - lot specific
• Conjugation: Unconjugated
• Storage: Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. 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.
• Stability: Expiration date is one (1) year from date of receipt.
• 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. Rockland Immunochemicals produces anti-epitope tag antibodies against many common epitope tags including Myc, GST, GFP, 6X His, MBP, FLAG™ and HA. Rockland Immunochemicals also produces antibodies to other tags including FITC, Rhodamine (TRITC), DNP and biotin.
• Synonyms: mouse anti-FLAG™ tag, Enterokinase Cleavage Site (ECS), mouse anti-DYKDDDDK, Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys
• Note: Anti-FLAG antibody has been tested by ELISA and western blot and is optimally suited for monitoring the expression of FLAG™ tagged fusion proteins. As such, this antibody can be used to identify fusion proteins containing the FLAG™ 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.