Goat anti-6-His Tag Antibody Affinity Purified
Product Details
Specifications
Request Formulation Change Phosphate Buffered Saline (PBS) containing 0.2% BSA and 0.05% Pro-Clean 400
Request Formulation Change Phosphate Buffered Saline (PBS) containing 0.2% BSA and 0.09% Sodium Azide
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Immunoglobulin concentration was determined using Beer’s Law where 1mg/mL IgG has an A280 of 1.4. Goats were immunized with 6-His (HHHHHH) conjugated to KLH. Antibody was isolated by affinity chromatography using the peptide immobilized on solid support and conjugated to horseradish peroxidase (HRP). This antibody does not detect all N-terminal 6-His tagged proteins.
Prior to conjugation, immunoglobulin concentration was determined using Beer’s Law where 1mg/mL IgG has an A280 of 1.4. Molar enzyme/antibody protein ratio is 4:1. Goats were immunized with 6-His (HHHHHH) conjugated to KLH.
Antibody was affinity purified using the peptide immobilized on solid support. Goat anti-6-His affinity purified antibodies were coupled to agarose beads using a cyanogen bromide method. Final concentration is 0.1 mg antibody in 0.4 ml of 50% slurry containing 0.2 ml agarose.
Immunoglobulin concentration was determined using Beer’s Law where 1mg/mL IgG has an A280 of 1.4, before coupling to agarose beads. Goats were immunized with 6-His (HHHHHH) conjugated to KLH. Antibody was isolated by affinity chromatography using the peptide immobilized on solid support and conjugated to fluorescein isothiocyanate (FITC). This antibody does not detect all N-terminal 6-His tagged proteins.
Prior to conjugation, immunoglobulin concentration was determined using Beer’s Law where 1mg/mL IgG has an A280 of 1.4.
Additional Product Information
The 6-His tag is a commonly used epitope tag that can be added to a recombinantly expressed protein for the purposes of protein purification or detection of protein. Typically utilizes metal affinity chromatography for purification and elutes well imidazole.
Applications
Not all listed applications have been specifically tested by our laboratory.
All western blot analysis is performed using 5% Milk-TBST for blocking and as antibody diluent. Primary antibody is incubated overnight.
Western blots are performed using Donkey anti-Goat IgG Heavy and Light Chain Antibody (Cat. No. A50-101P).