Thermo Fisher Scientific 6X His Tag Polyclonal Antibody, Biotin

Applications

Tested Dilution

Publications

Western Blot (WB)

1:500

-

Immunohistochemistry (IHC)

1:1,000-1:5,000

-

ELISA (ELISA)

1:10,000-1:50,000

-

Immunoprecipitation (IP)

1:2,000-1:10,000

-

Product Specifications

Species Reactivity

Tag

Host/Isotype

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

This antibody was purified from whole rabbit serum prepared by repeated immunizations with 6X His epitope tag peptide H-H-H-H-H-H conjugated to KLH using maleimide.

Conjugate

Biotin Biotin Biotin

Form

Lyophilized

Concentration

1 mg/mL

Purification

Affinity chromatography

Storage buffer

0.02M potassium phosphate, pH 7.2, with 0.15M NaCl, 10mg/mL BSA

Contains

0.01% sodium azide

Storage conditions

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Shipping conditions

Ambient (domestic); Wet ice (international)

Product Specific Information

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.

This affinity purified antibody is 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 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.

Target Information

The 6x His tag is a synthetic oligo peptide consisting of 6 consecutive histidine residues (HHHHHH). The His tag is commonly expressed as a tag at either N-or C-terminal regions of recombinant proteins to allow isolation or purification by immobilized metal affinity chromatography. Epitope tagging is a technique in which a known epitope is fused to a recombinant protein using genetic engineering. By choosing a particular epitope and recombinant protein combination, epitope tagging makes it possible to detect proteins for which no antibody is available. His epitope tagged proteins that contain a stretch of histidine residues at the carboxyl or amino terminus are used for purification of recombinant proteins by means of metal chelate chromatography due to metal binding properties of histidine clusters on His tagged proteins. For example, the affinity of the histidine-tag motif to Ni2+ by chelation is strong and selective enough to enable purification of the protein to homogeneity by affinity chromatography on a Ni2+-NTA adsorbent. His tag-specific antibodies are used to facilitate detection or coimmunopreciptation of tagged proteins.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.