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DYKDDDDK Antibody same epitope as Sigma's Anti-FLAG HRP Conjugated

Rabbit Polyclonal
NCI Collaboration
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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 1:1,000 for 60 min at RT.
Blocking:  MB-070 for 30 min at RT.
Predicted/Observed size:  49 kDa, 49 kDa for BAP FLAG.
Other band(s):  None.
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Number:
600-403-383
Size:
100 µg
Price:
$309.00
Shipping:
$40.00
Handling:
$20.00
Availability:
Ships next business day.

DYKDDDDK Antibody same epitope as Sigma's Anti-FLAG HRP Conjugated Properties

Description
Antibody for the detection of FLAG™ conjugated proteins (RABBIT) Antibody Peroxidase Conjugated - 600-403-383
Host
Rabbit
Known Cross Reactivity
Carboxy and amino terminal linked FLAG™ tagged recombinant proteins
Clonality
Polyclonal
Application
ELISA : 1:20,000
Western Blot : 1:2,000 - 1:5,000
Immunohistochemistry: 1:500 - 1:2,000
Other Dilution: User Optimized
Epitope Tag Type
FLAG
Physical State
Lyophilized
Shipping Condition
Ambient
Format
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) Gentamicin Sulfate. Do NOT add Sodium Azide!
Stabilizer
10 mg/mL Bovine Serum Albumin (BSA) - Immunoglobulin and Protease free
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DYKDDDDK Antibody same epitope as Sigma's Anti-FLAG HRP 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.
Synonyms
HRP
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.  Residues of glycine and cysteine were added to the carboxy terminal end to facilitate coupling.  This antibody reacts with FLAG conjugated proteins.
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 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.  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.
Purity/Specifity
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.
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 5199, Limerick, 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, 545-55. Fulton, J.E. et al., (1995) Functional analysis of avian class I (BFIV) glycoproteins by epitope tagging and mutagenesis in vitro. Eur J Immunol 25:7, 2069-76.
Specific Reference
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Product Type
Primary Antibodies;
Applications
ELISA, WB, IHC, IF, FC
Reacts With
human, mouse, rat, monkey
Catalog Number
200-301-268

Product Type
Secondary Antibodies;
Applications
ELISA, WB, IHC
Catalog Number
610-4302

Product Type
Secondary Antibodies;
Applications
ELISA, WB, IHC
Catalog Number
611-1302

Product Type
Supporting Reagents;
Applications
ELISA, FLISA, WB, IHC, IF, IP, FC
Catalog Number
B304

Product Label

 

Description
Peroxidase (Horseradish)
Color
Substrate dependent
Conjugation Reference
Modified from Farr & Nakane, J. Immunol. Methods 47; 129-144. 1981.
Molecular Weight
40000
Excitation Wavelength
Conjugation Chemistry
Periodate