House Secondary Banner
 
No Related Products Associated
GFP and Epitopes  >  Antibodies to FLAG™ and Antibodies to 6XHIS Tags
 

6X His Tag Antibody IRDye800® Conjugated

Mouse Monoclonal 33D10.D2 IgG1 kappa
Code

200-332-382
Size

100 µg
Price

$309.00
Availability

Ships next business day.
Order Offline:
Phone: 800.656.7625
E-mail: orders@rockland-inc.com
Fax: 610.367.7825
Bulk Orders:
Call or E-mail Us For Information
Fill out our Contact Form
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.
Shipping & Handling Fees Apply.

Shipping:
$40.00 to United States
Handling:
$20.00
Synonyms: anti-HIS, HIS Antibody, 6X His Tag Antibody, HHHHHH epitope tag antibody

6X His Tag Antibody IRDye800® Conjugated Properties

Description
Anti-6X HIS EPITOPE TAG (MOUSE) Monoclonal Antibody IRDye® 800 Conjugated - 200-332-382
Host
Mouse
Clonality
Monoclonal 33D10.D2 IgG1 kappa
Application
FLISA : 1:5,000 - 1:10,000
IF Microscopy : >1:5,000
Western Blot : 1:10,000 - 1:25,000
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
3.9 moles IRDye® 800 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
Download CofA
Lot#  

6X His Tag Antibody IRDye800® 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 biochemical properties of the tagged protein.  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
Fluorescence technology is widely used to detect proteins. However, many common visible fluorophores often result in considerable background fluorescence in the visible range. Visible fluorophores are rarely used for membrane-based protein detection because of this high background. IRDye® 800 and IRDye® 700DX antibody and reagent conjugates are specifically designed for protein detection methods that use longer-wavelength, near-infrared (IR) fluorophores to visualize proteins in western blotting and other applications. Very low background fluorescence in the IR range provides for a much higher signal-to-noise ratio than visible fluorophores. Detection levels in the picogram range on Western blots rival the sensitivity of chemiluminescence on film. IRDye® 800 conjugates are optimized for the Odyssey® Infrared Imaging System developed by LI-COR. IRDye® 800 conjugates are also suitable for immunofluorescence microscopy using commercially available excitation/emission filters in the 780nm/820nm range. Dual simultaneous labeling in western blots or microscopy is achieved when IRDye® 800 conjugates are used in conjunction with IRDye® 700DX or DyLight™680 conjugates. IRDye® 800 and IRDye® 700DX conjugates provide an ultra-sensitive and convenient alternative to standard chemiluminescent protein detection methods, as well as a valuable tool for multicolor imaging.
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
Log in to submit your product review.

No Related Products Associated

Product Label

 

Description
IRDye® 800
Color
Infra-Red
Conjugation Reference
Modified from LI-COR Biosciences, Lincoln, NE
Molecular Weight
1166
Excitation Wavelength
778
Conjugation Chemistry
N-hydroxysuccinimide (NHS) ester