Nickel Coated Plates Clear 8-well Strip
PRODUCT CODE: X-MTP-0001-5X
STORAGE: room temperature
Immobilized metal affinity interaction allows polyhistidine tag containing protein to be captured on surfaces that contain chelated divalent ions such as Ni2+, Cu2+, Co2+ and Zn2+. Cobalt in comparison to Nickel has lower affinity but highly specific binding.
BioThinx Nickel coated plates are designed for capture, detection and purification of polyhistidine tagged proteins and peptides.
This product may be used with His-tagged molecules in various applications such as recombinant protein expression screening, immunoabsorbtion assays, biochemical assays, competition assays and protein purification. The captured proteins or peptides can be detected using standard ELISA techniques or eluted for further analysis.
PRECAUTIONS AND DISCLAIMER
This product is for LABORATORY RESEARCH USE ONLY, not for diagnostic, therapeutic, drug, household, or other uses. Please consult the Material Safety Data Sheet for information regarding hazards and safe handling practices.
Nickel coated micro assay plate: 96 wells, configured in twelve 1×8 strips, each coated plate is packed in a barrier bag with desiccant. The wells are coated to a 100μl depth and are supplied pre-blocked.
PREPARTION AND HANDLING
The following protocol is a simple direct ELISA protocol and the protocol and reagents used will have to be optimized for specific applications and assays. Avoid using buffer that contain EDTA or other metal chelators, avoid reducing agents and avoid imidazole.
1. Wash the wells to be used with 200μl Wash Buffer (tris buffered saline or phosphate buffered saline, pH 7-7.5, containing 0-05 % TWEEN® 20 or an appropriate Wash Buffer of choice.
2. Dilute the sample with (tris buffered saline or phosphate buffered saline, pH 7-7.5, containing 0.05 % BSA or an appropriate Dilution Buffer of choice and add 100μl diluted sample to each well.
3. Incubate at room temperature for 1-2 hours with shaking.
4. Wash each well three times with 200μl Wash Buffer.
5. Add 100μl enzyme labelled primary antibody.
6. Incubate at room temperature for 0.5-1 hour with shaking.
7. Wash each well three times with 200μl Wash Buffer.
8. Detect the label signal with appropriate substrate.
STORAGE / STABILITY
Store unopened at ambient temperature. Once opened the plates can be stored in the resealable bag (ZipLoc) with desiccant.
RECOMMENDED RETEST DATE
1. Hochuli, E., et al., New metal chelate adsorbent for proteins and peptides containing neighbouring histidine residues, J. Chromatogr., 411, 177-84 (1987).
2. Block, H., et al., Immobilized-metal affinity chromatography (IMAC): a review. In: Richard, B.R. and Deutscher, M.P., eds. Methods Enzymol. 463, 439–73 (2009).