6xHistidine Epitope Tag (HHHHHH) Rabbit Polyclonal Antibody

CAT#: DP3514

6xHistidine Epitope Tag (HHHHHH) rabbit polyclonal antibody, Aff - Purified

Product Images

Western blot (ECL) detection of different His-tagged proteins. For Western blot analysis each lane was loaded with 12, 5-50 ng with the following His-tagged proteins: Lane 1: TbTX, Lane 2: IL-3, Lane 3: VEGF121, Lane 4: PlGF-1, Lane 5: Lac I repressor, Lane

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  Western blot (AP) of several His-tagged proteins from supernatants (A) and after purification (B) by affinity chromatography using metal chelate columns (Ni2+, Co2+) as matrix. The anti-His-tag antibody concentration used was 0.1g/ml.

Specifications

Product Data

• Applications: WB
• Recommended Dilution: Western blot (0.5-1 µg/ml): The antibody was tested using several His-tagged proteins expressed in insect cells and E. coli.
• Host: Rabbit
• Clonality: Polyclonal
• Immunogen: Different highly purified 6x His-tagged proteins (C-terminal) produced in insect cells.
• Specificity: Western analysis with several His-tagged proteins either expressed in insect cells or E.coli showed that the antibody recognizes all tested proteins fused to a C-terminal but not to a N-terminal His-tag.
  The antibody might be a very good tool to test supernatants or cell lysates for expression of recombinant proteins.
  The anti-His-Tag antibody is able to detect recombinant proteins in the conditioned media from insect cells and total lysate from E.coli.
• Formulation: PBS, pH 7.4, without preservatives.
  State: Aff - Purified
  State: Lyophilized purified IgG fraction.
• Reconstitution Method: Restore in sterile water to a concentration of > 0.5 mg/ml.
• Purification: Antigen Affinity Chromatography using a His-peptide as matrix.
• Conjugation: Unconjugated
• Storage: The lyophilized IgG is stable at 2-8°C for one month from despatch and for one year when kept at -20°C.
  The reconstituted antibody can be stored at 2-8°C for one month or at -20°C for one year.
  Avoid repeated freezing and thawing.
• Background: In the last couple of years many peptide sequences/epitopes for the purification of recombinant proteins have been established. These so-called “tags” can be used e.g. to determine the cellular localization or to quantify proteins. The polyhistidine “tag” (His-tag) is the most used affinity epitope for the purification of recombinant proteins [1]. Proteins with a polyhistidine tag (e.g. 6xHis or 8xHis) can be purified in one step using a metal-chelate column (e.g. Ni2+, Zn2+, Cu2+ or Co2+) and imidazole as eluent. This method now is a very attractive system for the purification of larger amounts proteins for structural and functional studies. So far His-tagged proteins were successfully purified from different expression systems like E. coli, yeast, insect cells and plant cells [1,2]. An important requirement beside the efficient and robust purification method is the availability of a fast detection system for checking the purification steps of these His-tagged proteins if no specific antibody is available.