Anti-FLAG-conjugated proteins, Fluorescein conjugated, clone 29E4.G7

Art Nr.: 200-302-383, Rockland
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DYKDDDDK Antibody same epitope as Sigmas Anti-FLAG Fluorescein Conjugated Properties



Description Antibody for the detection of FLAG™ conjugated proteins (MOUSE) Monoclonal Antibody Fluorescein Conjugated - 200-302-383

Host Mouse

Known Cross Reactivity Carboxy and amino terminal linked FLAG™ tagged recombinant proteins

Clonality Monoclonal 29E4.G7 IgG2a kappa

Application IF Microscopy : 1:500 - 1:2,500
Other Dilution: User Optimized

Epitope Tag Type FLAG

Physical State Lyophilized

Shipping condition Ambient

Format IgG

Label Fluorescein (FITC)

FP Value 3.1 moles Fluorescein (FITC) per mole of IgG

Concentration 1.0 mg/mL by UV absorbance at 280 nm

Buffer 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

Reconstitution Volume 100 µL

Reconstitution Buffer Restore with deionized water (or equivalent)

Preservative 0.01% (w/v) Sodium Azide

Stabilizer 10 mg/mL Bovine Serum Albumin (BSA) - Immunoglobulin and Protease free  

DYKDDDDK Antibody same epitope as Sigmas Anti-FLAG Fluorescein Conjugated Description

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.

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. Residues of glycine and cysteine were added to the termini to facilitate coupling.

Immunogen Type

Peptide

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. 

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 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.

Purity/Specifity

This antibody is directed against the FLAG? epitope tag and is useful in determining its presence in over expressed proteins in various assays. The antibody recognizes the FLAG? epitope tag (Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys) fused to either the amino- or carboxy- termini of targeted proteins in transfected or transformed cells.

Disclaimer Note-General

This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 326, Gilbertsville, Pennsylvania, USA.

General Reference Chubet, R.G. and Brizzard, B.L. (1996) Vectors for expression and secretion of FLAG epitope-tagged proteins in mammalian cells. Biotechniques 20(1):136-141. Slootstra, J.W., et al. (1997) Identification of new tag sequences with differential and selective recognition properties for the anti-FLAG monoclonal antibodies M1, M2 and M5. Mol. Divers. 2(3):156-164. Robeva, A.S., et al. (1996) Double tagging recombinant A1- and A2A-adenosine receptors with hexahistidine and the FLAG epitope. Development of an efficient generic protein purification procedure. J. Biochem. Pharmacol. 51(4):545-555.

Specific Reference

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