Key Product Details

Species Reactivity

Epitope Tag

Applications

Dot Blot, ELISA, Fluorophore-linked immunosorbent assay, Immunocytochemistry/ Immunofluorescence, Western Blot

Label

DyLight 488 (Excitation = 493 nm, Emission = 518 nm)

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # 33D10.D2.G8

Concentration

LYOPH mg/ml

View all His Tag Antibodies »

Product Specifications

Immunogen

His Tag Antibody (33D10.D2.G8) was produced in mice by repeated immunizations with 6X His epitope tag peptide H-H-H-H-H-H conjugated to KLH using maleimide.

Reactivity Notes

6x His tag specific

Specificity

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.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

Description

Store vial at 4C prior to restoration. For extended storage aliquot contents and freeze at -20C 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 4C as an undiluted liquid. Dilute only prior to immediate use.

This protein-A purified antibody is directed against the 6X His motif and is useful in determining its presence in various assays

Scientific Data Images for His Tag Antibody (33D10.D2.G8) [DyLight 488]

Western Blot: His Tag Antibody (33D10.D2.G8) [DyLight 488] [NBP1-97329] - Western Blot of His Tag antibody (33D10.D2.G8) [DyLight 488].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-His Tag antibody at 1:5,000 overnight at 4C.Secondary antibody: Peroxidase mouse secondary antibody at 1:20,000 for 30 min at RT.Block: 5% BLOTTO for 1 hr at RT.
Immunocytochemistry/Immunofluorescence: His Tag Antibody (33D10.D2.G8) [DyLight 488] [NBP1-97329] - DyLight(TM) dyes can be used for multi-color immunofluorescence microscopy with uniform fluorescence intensity throughout the image. DyLight(TM) dyes are exceptionally bright and photostable and are optimized for microscopy and microarray detection methods. This image shows anti-histone detection using a DyLight(TM) 488 conjugate (green). Anti-Tubulin was detected using a DyLight(TM) 549 conjugate (red). Nuclei were counter-stained using DAPI (blue). The image was captured using an Axio Imager.Z1 (Zeiss Micro Imaging Inc).
Immunocytochemistry/Immunofluorescence: His Tag Antibody (33D10.D2.G8) [DyLight 488] [NBP1-97329]
View 2 more images

Applications for His Tag Antibody (33D10.D2.G8) [DyLight 488]

Application
Recommended Usage

Fluorophore-linked immunosorbent assay

>1:20000

Immunocytochemistry/ Immunofluorescence

>1:5000

Western Blot

1:10000 - 1:25000
Application Notes
This product has been tested by ELISA and western blot. The emission spectra for this DyLight(TM) conjugate match the principle output wavelengths of most common fluorescence instrumentation.

Formulation, Preparation, and Storage

Purification

Protein A purified

Reconstitution

Reconstitute with 100 ul deionized water (or equivalent)

Formulation

Lyophilized from 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 10 mg/mL Bovine Serum Albumin (BSA) - Immunoglobulin and Protease free

Preservative

0.01% Sodium Azide

Concentration

LYOPH mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store lyophilized antibody at 4C in the dark. Aliquot reconstituted liquid and store at -20C. Avoid freeze-thaw cycles.

Background: His Tag

A his-tag (also known as histidine tag or polyhistidine tag) is a common epitope tag that typically consists of at least 6 histidine residues fused to either the carboxyl (C-) or amino (N-) terminus of a targeted recombinant protein to facilitate its purification and detection (1). The most common his-tag is the hexahistidine (His6/6-His) tag which has a theoretical molecular weight of 0.8kda (1). The histidine residues readily interact with transition metal ions such as Co2+, Ni2+, Cu2+, and Zn2+, making immobilized metal-affinity chromatography (IMAC) the preferred technique for his-tag purification (1, 2). Metal ions are immobilized and bound to by His-tags in the IMAC column via the histidine imidazole ring. The tagged protein can be eluted off the column by washing with buffers containing a low concentration of imidazole (1, 2). Due to its relatively small size, low immunogenicity, versatility under denaturing conditions, and minimal interference with the structure and function of proteins, the his-tag is one of the most widely used tags for protein purification (1-3).

References

1. Malhotra, A. (2009). Tagging for protein expression. Methods in Enzymology, Guide to Protein Purification, 2nd Edition, 463, 239-258. https://doi.org/10.1016/s0076-6879(09)63016-0

2. Terpe, K. (2003). Overview of tag protein fusions: from molecular and biochemical fundamentals to commercial systems. Applied Microbiology and Biotechnology, 60(5), 523-533. https://doi.org/10.1007/s00253-002-1158-6

3. Booth, W. T., Schlachter, C. R., Pote, S., Ussin, N., Mank, N. J., Klapper, V., ... Chruszcz, M. (2018). Impact of an N-terminal polyhistidine tag on protein thermal stability. ACS Omega, 3(1), 760-768. https://doi.org/10.1021/acsomega.7b01598

Long Name

Histidine Tag

Alternate Names

polyHistidine

Additional His Tag Products

Product Documents for His Tag Antibody (33D10.D2.G8) [DyLight 488]

Product Datasheets

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

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Product Specific Notices for His Tag Antibody (33D10.D2.G8) [DyLight 488]



DyLight (R) is a trademark of Thermo Fisher Scientific Inc. and its subsidiaries.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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