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GFP and Epitopes  >  Antibodies to FLAG™ and Antibodies to 6XHIS Tags
 

6X His Tag Antibody Biotin Conjugated

Mouse Monoclonal 33D10.D2 IgG1 kappa
Code

200-306-382
Size

100 µg
Price

$309.00
Availability

Ships next business day.
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ELISA of Mouse anti-6xHIS Tag Antibody.
Antigen: HIS-tagged purified protein and E. coli cell lysates expressing HIS-Tagged construct, GST- and RON-tagged purified proteins.
Coating amount: 0.15ug per welll.
Primary antibody: 6xHIS Tag antibody at 100ug/mL.
Dilution series: 2-fold.
Mid-point concentration: 200ng/mL.
Secondary antibody: Peroxidase mouse secondary antibody at 1:10,000.
Substrate: TMB (p/n TMBE-1000).
Western Blot of Mouse anti-6xHIS Tag Antibody.
Lane 1: 100ng Purified histidine-tagged recombinant protein.
Lane 2: 200ng E. coli cell lysate containing histidine-tagged expression construct.
Lane 3: 100ng Purified GST-tagged recombinant protein.
Lane 4: 100ng Purified FLAG-tagged recombinant protein.
Primary antibody: Mouse anti-6xHIS Tag antibody at 1:5,000 overnight at 4°C.
Secondary antibody: Peroxidase mouse secondary antibody at 1:20,000 for 30 min at RT.
Block: 5% BLOTTO for 1 hr at RT.
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Synonyms: anti-HIS, HIS Antibody, 6X His Tag Antibody, HHHHHH epitope tag antibody

6X His Tag Antibody Biotin Conjugated Properties

Description
Anti-6X HIS EPITOPE TAG (MOUSE) Monoclonal Antibody Biotin Conjugated - 200-306-382
Host
Mouse
Clonality
Monoclonal 33D10.D2 IgG1 kappa
Application
ELISA : 1:20,000 - 1:100,000
Western Blot : 1:2,000 - 1:10,000
Immunochemistry: 1:1,000 - 1:5,000
Other Dilution: FLUORESCENT POLARIZATION: 2 - 20 nMFRET: 2 - 20 nM
Epitope Tag Type
6X HIS
Physical State
Lyophilized
Shipping condition
Ambient
Format
IgG
Label
FP Value
10-20 moles Biotin 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
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6X His Tag Antibody Biotin 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 6X His epitope tag peptide H-H-H-H-H-H conjugated to KLH using maleimide.
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
Anti-6X His is optimally suited for monitoring expression of His-tagged fusion proteins. As such, anti-6X His/6X His can be used to identify fusion proteins containing the 6X His epitope.  The antibody recognizes the His tag fused either to the amino- or carboxy- termini of targeted proteins.  This antibody has been tested by ELISA and western blotting against both the immunizing peptide and His-containing recombinant proteins.  Although not tested, this antibody is likely functional for immunoprecipitation and immunocytochemistry.
Purity/Specifity
This protein-A purified antibody is directed against the 6X His motif and is useful in determining its presence in various assays. This monoclonal anti-6X His tag antibody detects over-expressed proteins containing the 6X His epitope tag.  To date, this antibody has reacted with all His tagged proteins so far tested.  In western blotting of bacterial extracts, the antibody does not cross-react with endogenous proteins.  The antibody recognizes the His-tag (His-His-His-His-His-His) 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
Specific Reference
Brandsma, M ; X Wang, H Diao, SE. Kohalmi1, AM Jevnikar and S. Ma (2009) A Proficient Approach to the Production of Therapeutic Glucagon-Like Peptide-1 (GLP-1) in Transgenic PlantsThe Open Biotechnology Journal, 3, 57-66 57 1874-0707/09
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Product Label

 

Description
Biotin
Color
n/a
Conjugation Reference
Modified from Bayer & Wilchek Methods in Enzymology 184; 138-160, 1990.
Molecular Weight
455
Excitation Wavelength
Conjugation Chemistry
N-hydroxysuccinimide (NHS) ester