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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 biochemical properties of the tagged protein.  Most often, sequences encoding the epitope tag are included with the 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.

Product Details

Antibody for the detection of FLAG™ conjugated proteins (RABBIT) - 600-401-383
rabbit antibody for the detection of FLAG™ conjugated proteins, rabbit anti DYKDDDDK
Rabbit
Polyclonal
IgG

Target Details

FLAG-Tag (C-Term)
Conjugated Peptide
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.
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.  In western blotting of bacterial extracts, the antibody does not cross-react with endogenous proteins.

Application Details

ELISA, WB
IF, IHC, IP, Multiplex  - View References
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.  This antibody has been tested by ELISA and western blotting against both the immunizing peptide and FLAGä containing recombinant proteins.  Although not tested, this antibody is likely functional for immunoprecipitation, 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.

Formulation

Liquid (sterile filtered)
1.17mg/mL by UV absorbance at 280 nm
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
0.01% (w/v) Sodium Azide
None

Shipping & Handling

Dry Ice
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.
Expiration date is one (1) year from date of receipt.
(). Mediobasal hypothalamic FKBP51 acts as a molecular switch linking autophagy to whole-body metabolism. Science Advances
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IP, Co-IP
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WB, IB, PCA
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WB, IB, PCA
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WB, IB, PCA
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IHC, ICC, Histology; WB, IB, PCA
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Undefined
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IHC, ICC, Histology; WB, IB, PCA
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WB, IB, PCA
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WB, IB, PCA
(). Cross Talk between ARF1 and RhoA Coordinates the Formation of Cytoskeletal Scaffolds during Chlamydia Infection. mBio.
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WB, IB, PCA
(). Rab32 uses its effector reticulon 3L to trigger autophagic degradation of mitochondria-associated membrane (MAM) proteins. Biol Direct.
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WB, IB, PCA
(). Modulation of virus-induced NF-κB signaling by NEMO coiled coil mimics. Nat Commun.
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IP, Co-IP
(). LITESEC-T3SS-Light-controlled protein delivery into eukaryotic cells with high spatial and temporal resolution. Nat Commun.
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WB, IB, PCA
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WB, IB, PCA
(). Pathogenic mutations reveal a role of RECQ4 in mitochondrial RNA: DNA hybrid formation and resolution. Sci Rep.
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IP, Co-IP; WB, IB, PCA
(). Mechanosensitive collaboration between integrins and connexins allows nutrient and antioxidant transport into the lens. J Cell Biol.
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Undefined
(). CdbA is a DNA-binding protein and c-di-GMP receptor important for nucleoid organization and segregation in Myxococcus xanthus. Nature Communications
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WB, IB, PCA
(). Heat shock protein B1 is a key mediator of prolactin-induced beta-cell cytoprotection against oxidative stress. Free Radic Biol Med.
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WB, IB, PCA
(). Regulation of somatostatin receptor 2 trafficking by C-tail motifs and the retromer. Endocrinology.
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IF, Confocal Microscopy; WB, IB, PCA; Multiplex Assay
(). Epigenetic derepression of FKBP5 by aging and stress contributes to NF-ĸB-driven inflammation and cardiovascular risk. Proc Natl Acad Sci USA
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WB, IB, PCA
(). Robust Antibacterial Activity of Tungsten Oxide (WO3-x) Nanodots. Chemical Research in Toxicology
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(). Resolving the Combinatorial Complexity of Smad Protein Complex Formation and Its Link to Gene Expression. Cell Systems
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IP, Co-IP
(). Evolutionary divergence of the sex-determining gene MID uncoupled from the transition to anisogamy in volvocine algae. Development
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WB, IB, PCA
(). Construction of a tri-chromatic reporter cell line for the rapid and simple screening of splice-switching oligonucleotides targeting DMD exon 51 using high content screening. PLOS One
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WB, IB, PCA
(). SARS-Coronavirus Open Reading Frame-3a drives multimodal necrotic cell death. Cell Death & Disease
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(). Protease-activated receptor-4 and P2Y12 Dimerize, Co-Internalize and Activate Akt Signaling via Endosomal Recruitment of β-arrestin. Journal of Biological Chemistry
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IF, Confocal Microscopy; IP, Co-IP; WB, IB, PCA
(). Connexin 50 Functions as an Adhesive Molecule and Promotes Lens Cell Differentiation. Scientific Reports
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WB, IB, PCA
(). De novo mutations in EBF3 cause a neurodevelopmental syndrome. Am J Hum Genet.
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WB, IB, PCA
(). Recycling and Endosomal Sorting of Protease-activated Receptor-1 Is Distinctly Regulated by Rab11A and Rab11B Proteins. Journal of Biological Chemistry
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E, EIA
(). S6K-STING interaction regulates cytosolic DNA-mediated activation of the transcription factor IRF3. Nature Immunology
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WB, IB, PCA
(). Protease-activated Receptor-4 Signaling and Trafficking Is Regulated by the Clathrin Adaptor Protein Complex-2 Independent of β-Arrestins. Journal of Biological Chemistry
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IF, Confocal Microscopy
(). FKBP51 inhibits GSK3β and augments the effects of distinct psychotropic medications. Mol Psychiatry.
Applications
IP, Co-IP; WB, IB, PCA
(). GPCR sorting at multivesicular endosomes. Methods Cell Biol.
Applications
IF, Confocal Microscopy
(). Chaperoning epigenetics: FKBP51 decreases the activity of DNMT1 and mediates epigenetic effects of the antidepressant paroxetine. Sci Signal.
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WB, IB, PCA
(). Interaction with the effector dynamin-related protein 1 (Drp1) is an ancient function of Rab32 subfamily proteins. Cell Logist.
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IP, Co-IP
(). Essential role of the chaperonin CCT in rod outer segment biogenesis. Invest Opthalmol. Vis Sci.
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WB, IB, PCA
(). A novel COX-independent mechanism of sulindac sulfide involves cleavage of epithelial cell adhesion molecule protein. Exp Cell Res.
Applications
IF, Confocal Microscopy; Multiplex Assay
(). Mi-2/NuRD chromatin remodeling complexes regulate B and T-lymphocyte development and function. Immunol Rev.
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WB, IB, PCA
(). Association of FKBP51 with priming of autophagy pathways and mediation of antidepressant treatment response: evidence in cells, mice, and humans. PLoS Med.
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WB, IB, PCA
(). Splice isoforms of phosducin-like protein control the expression of heterotrimeric G proteins. J Biol Chem.
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WB, IB, PCA
(). Phosphorylation of the adaptor ASC acts as a molecular switch that controls the formation of speck-like aggregates and inflammasome activity. Nat Immunol.
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WB, IB, PCA
(). Loss of the methyl lysine effector protein PHF20 impacts the expression of genes regulated by the lysine acetyltransferase MOF. Biol Chem.
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WB, IB, PCA
(). Metal allergens nickel and cobalt facilitate TLR4 homodimerization independently of MD2. EMBO Rep.
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WB, IB, PCA
(). Transgenic expression of human heme oxygenase‐1 in pigs confers resistance against xenograft rejection during ex vivo perfusion of porcine kidneys. Xenotransplantation.
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WB, IB, PCA
(). Y3IP1, a nucleus-encoded thylakoid protein, cooperates with the plastid-encoded Ycf3 protein in photosystem I assembly of tobacco and Arabidopsis. Plant Cell.
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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 5199, Limerick, Pennsylvania, USA.