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Antibody for the detection of FLAG™ conjugated proteins (RABBIT) Antibody Biotin Conjugated
DYKDDDDK Antibody same epitope as Sigma's Anti-FLAG Biotin Conjugated
- SPECIFICATION
- CITATIONS
- PROTOCOLS
- BACKGROUND
| Host | Rabbit |
|---|---|
| Conjugate | Biotin |
| FP Value | 10-20 |
| Clonality | Polyclonal |
Application 
| WB, IHC, E, I, LCI |
| Application Note | This antibody 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 the carboxy or amino- terminus of targeted proteins. This antibody has been tested by ELISA and western blotting against both the immunizing peptide and FLAG™ 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. Rockland Immunochemical's polyclonal antibody to detect FLAG™ conjugated proteins binds FLAG™ 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. |
| Physical State | Lyophilized |
| Buffer | 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2 |
| Immunogen | This antibody was purified from whole rabbit serum prepared by repeated immunizations with the Enterokinase Cleavage Site (ECS) peptide DYKDDDDK (Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys) conjugated to KLH using maleimide. This antibody reacts with FLAG conjugated proteins. |
| Reconstitution Volume | 100 µL |
| Reconstitution Buffer | Restore with deionized water (or equivalent) |
| Stabilizer | 10 mg/mL Bovine Serum Albumin (BSA) - Immunoglobulin and Protease free |
| Preservative | 0.01% (w/v) Sodium Azide |
| Purity | This affinity purified antibody is directed against the FLAG™ motif and is useful in determining its presence in various assays. This polyclonal anti-FLAG™ tag antibody detects over-expressed proteins containing the FLAG™ epitope tag. To date this antibody has reacted with all amino- terminal FLAG™ tagged proteins so far tested. In western blotting of bacterial extracts the antibody does not cross-react with endogenous proteins. |
|---|---|
| Storage Condition | 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. |
| Precautions Note | This product is for research use only and is not intended for therapeutic or diagnostic applications. |
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Provided below are standard protocols that you may find useful for product applications.
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.
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