Description
Anti-His Affinity Gel is prepared by conjugating high quality murine IgG1 monoclonal antibody with Sepharose 4B gel. This product has a high His label fusion protein loading capacity (at least 1.1 mg protein/mL gel) and less non-specific binding of miscellaneous proteins. It can be used for immunoprecipitation (IP) and purification of fusion proteins with His labels.
Features
- Specific—highly specific monoclonal antibody
- High yield and high purity
- Versatile—It can be used for protein purification or COIP
- Convenient—reagents and peptides to elute are available separately
Applications
- Immunoprecipitation (IP)
- Purification of fusion proteins with His labels.
Specifications
| Isotype | Mouse IgG1 |
| Purity | Protein A |
| Antibody Concentration | 7.5 g antibody/L Gel |
| Protein binding capacity | ≥1.1 mg protein/mL Gel |
| Storage buffer | TBS, 50% glycerin, pH7.4, contains 0.02% (w/v) sodium azide |
Components
| Components No. | Name | 20589ES03 | 20589ES08 | 20589ES25 | 20589ES60 |
| 20589 | Anti-His Affinity Gel | 1 mL | 5 mL | 25 mL | 100 mL |
Storage
The unopened products should be stably stored at -25℃ ~ -15℃ for 1 year. Do not freeze in solution without glycerin!
This gel captures all common polyhistidine-tag formats: 6×His, 8×His, 10×His, and other His-tag lengths. The anti-His monoclonal antibody (IgG1, 7.5 g/L gel) recognizes the His-tag epitope regardless of the number of histidine residues (within reason). It provides an antibody-based alternative to Ni²⁺-NTA resins, which can be problematic with proteins sensitive to nickel or in the presence of chelating agents.
Key differences: (1) No nickel required — avoids issues with nickel toxicity for downstream assays and proteins sensitive to metal chelation; (2) Elution with imidazole is NOT required — elute with low-pH glycine or His peptide; (3) Works in the presence of EDTA and chelating agents; (4) Lower non-specific binding from histidine-rich endogenous proteins that can contaminate Ni-NTA eluates.
Use standard protein binding conditions: equilibrate in TBS (pH 7.4) or HEPES buffer. Add clarified, filtered lysate (0.22 µm filtered) and incubate 1–4 h at 4°C with gentle rotation. Avoid high imidazole in binding buffer (unlike Ni-NTA). Wash with TBS + 10–20 mM imidazole is NOT required; standard TBS wash is sufficient.
Preferred elution: His peptide (1 mg/mL in TBS) for native, functional protein. Alternatively: low-pH glycine buffer (50–100 mM, pH 2.5–3.0); neutralize the eluate immediately with 1 M Tris-HCl pH 9.0. Unlike Ni-NTA, high-imidazole elution is not the standard method for this antibody-based gel.
The antibody-based gel requires the His-tag epitope to be accessible. In denaturing conditions (8 M urea, 6 M guanidine-HCl), epitope accessibility can vary. For inclusion body purification under denaturing conditions, Ni-NTA resin is typically more robust. Validate binding under your specific denaturing conditions before use.
The following customization and add-on services may be available for this product type through the supplier. For inquiries and pricing, contact support@biohippo.com.
Customization Options
- Bulk Quantities: Larger volumes and custom pack sizes beyond the standard catalog may be available for high-throughput core facilities and industrial customers. Contact us to inquire about bulk pricing.
- Custom Antibody Coupling (Magnetic Beads): Custom magnetic beads conjugated to your specific antibody or protein ligand may be available through Yeasen's OEM and custom services program. These services leverage the same bead platform used for catalog products.
- OEM Manufacturing: Custom bead formulations for diagnostic device development, including magnetic beads for nucleic acid isolation, IP-grade beads, and affinity capture beads, may be available at manufacturing scale.
To inquire about custom options or bulk pricing, contact support@biohippo.com.
