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GFP and Epitopes  >  Sample Size Primary Antibodies
 

6X His Tag Antibody

Mouse Monoclonal 33D10.D2 IgG1 kappa
Code

200-301-382S
Size

25 µL
Price

$95.00
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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Shipping:
$85.00 to United States
Handling:
$20.00
Synonyms: anti-HIS, HIS Antibody, 6X His Tag Antibody, HHHHHH epitope tag antibody

6X His Tag Antibody Properties

Description
Anti-6X HIS EPITOPE TAG (MOUSE) Monoclonal Antibody - 200-301-382S
Host
Mouse
Clonality
Monoclonal 33D10.D2 IgG1 kappa
Application
ELISA : 1:20,000 - 1:100,000
FlowCytometry : User Optimized
Western Blot : 1:2,000 - 1:10,000
Immunochemistry: User Optimized
ImmunoPrecipitation: 1:200
Other Dilution: User Optimized
Epitope Tag Type
6X HIS
Physical State
Liquid (sterile filtered)
Shipping condition
Dry Ice
Format
IgG1
Label
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
Preservative
0.01% (w/v) Sodium Azide
Stabilizer
None
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6X His Tag Antibody Description

Background
6X His antibody is an Epitope tag which is a short peptide sequence that is 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.
Immunogen
This His 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 6X His Tag Antibody at -20° C or below prior to opening. This vial contains a relatively low volume of reagent (25 µL). To minimize loss of volume dilute 1:10 by adding 225 µL of the buffer stated above directly to the vial. Recap, mix thoroughly and briefly centrifuge to collect the volume at the bottom of the vial. Use this intermediate dilution when calculating final dilutions as recommended below. Store the vial at -20°C or below after dilution. Avoid cycles of freezing and thawing.
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 that contain 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
6X His antibody is affinity purified, directed against the 6X His motif and is useful in determining its presence in various assays. This polyclonal 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 5199, Limerick, Pennsylvania, USA.
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