6xHistidine Epitope Tag (HHHHHH) Chicken Polyclonal Antibody

CAT#: AP31792PU-N

6xHistidine Epitope Tag (HHHHHH) chicken polyclonal antibody, Purified

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Specifications

Product Data

• Applications: IP, WB
• Recommended Dilution: Western Blot.
  Recommended Dilutions: Use 1/5000-1/10000 as starting points for western blotting.
• Host: Chicken
• Isotype: Ig
• Clonality: Polyclonal
• Immunogen: 6X-Histidine peptide conjugated to Keyhole Limpet Hemocyanin protein and emulsified in Freund’s adjuvants.
  After multiple injections, eggs were collected from the hens, IgY fractions were prepared from the yolks, and then affinity-purified antibodies were prepared using 6X-Histidine peptide conjugated to an agarose matrix.
  To make up the final mixture, the IgY fraction was spiked with affinity purified antibody, mixed with Glycerol and Thimerosal, and filter-sterilized.
• Specificity: This antibody recognizes the 6xHIS-Tag with high titre and high specificity in Western Blot.
  Quality Control: This antibody preparation was analyzed by Western blotting (at a dilution of 1/10000) using tissue extracts expressing a recombinant protein tagged with the 6X-HIS marker. Secondary antibodies were HRP-labeled affinity-purified Goat anti-Chicken IgY (1/5000 dilution).
• Formulation: Sodium Phosphate-Buffered (10 mM) isotonic (0.9%, w/v) Saline (PBS, pH 7.2) (50% by volume) with 50% Glycerol as an anti-freezing agent and 0.01% Thimerosal as preservative.
  State: Purified
  State: Mixture of IgY and Affinity-purified fractions
• Concentration: 10 mg/ml (based on absorbance at 280 nm)
• 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. 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. 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.
• Synonyms: 6xHis-Tag, His Tag, HIS6 Tag, HHHHHH Tag