DYKDDDDK Tag (FLAG) Antibody Peroxidase Conjugated

SKU: orb345855

Description

DYKDDDDK antibody (Peroxidase)

Images & Validation

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Item 1 of 2

Tested Applications	ELISA, IHC, WB
Dilution Range	ELISA: 1:20,000, IHC: 1:500 - 1:2,000, WB: 1:2,000 - 1:5,000
Application Notes	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 amino- and carboxy- termini 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. Biorbyt'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. Read more...

Key Properties

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Antibody Type	Primary Antibody
Host	Rabbit
Clonality	Polyclonal
Isotype	IgG
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.
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 at either the N or C terminus. In western blotting of bacterial extracts the antibody does not cross-react with endogenous proteins.
Conjugation	HRP

Storage & Handling

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Storage	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.
Form/Appearance	Lyophilized
Buffer/Preservatives	Preservative: 0.01% (w/v) Gentamicin Sulfate. Do NOT add Sodium Azide!. Stabilizer: 10 mg/mL Bovine Serum Albumin (rAlbumin) - Immunoglobulin and Protease free; Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Concentration	1.0 mg/mL
Expiration Date	12 months from date of receipt.
Disclaimer	For research use only

Alternative Names

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rabbit anti-FLAG antibody peroxidase conjugation, HRP conjugated rabbit anti-FLAG antibody, anti-Flag tagged, anti-DYKDDDDK

Quality Guarantee

Explore bioreagents carefree to elevate your research. All our products are rigorously tested for performance. If a product does not perform as described on its datasheet, our scientific support team will provide expert troubleshooting, a prompt replacement, or a refund. For full details, please see our Terms & Conditions and Buying Guide. Contact us at support@biorbyt.com.

DYKDDDDK Tag (FLAG) Antibody Peroxidase Conjugated

Western blot of Peroxidase conjugated Rabbit Anti-DYKDDDDK same epitope as Sigma's Anti-FLAG antibody. Lane 1: BAP FLAG C-Term. Lane 2: BAP FLAG N-Term. Load: 50 ng per lane. Primary antibody: None. Secondary antibody: Peroxidase rabbit secondary antibody at 1:1000 for 60 min at RT. Blocking: orb348637 for 30 min at RT. Predicted/Observed size: 49 kDa, 49 kDa for BAP FLAG. Other band(s): None.

Western Blot of Peroxidase conjugated Rabbit Anti-DYKDDDDK same epitope as Sigma's Anti-FLAG antibody. Lane 1: Opal Presatained Molecular Weight Marker. Lane 2: FLAG Positive Control Lysate Reduced 5 µg [+]. Lane 3: 12 Epitope Tag Lysate-GST Reduced 5 µg [+]. Lane 4: MBP Reduced 0.1 µg [-]. Lane 5: rGST Reduced 0.1 µg [-]. Primary Antibody: Antibody for the detection of FLAG™ HRP 1.0 µg/ml overnight at 2-8°C. Secondary Antibody: Goat Anti-Rabbit IgG HRP MX10 (p/n orb347654) at 1:70000 for 30 mins at RT. Block: Universal Block Out Buffer (p/n orb348644) 1hr RT. Exposure 1sec.

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Protocol Information

Western Blot (IB, immunoblot)

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IHC

Immunohistochemistry

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ELISA

Enzyme-linked Immunosorbent Assay (EIA)

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