HIS-Select^® Plates

• iLAP^® Plates
• HIS-Select^® High Capacity (HC) Nickel Coated Plates
• HIS-Select^® High Sensitivity (HS) Nickel Coated Plates
• HIS-Select^® Filter Plates

Integrated Lysis and Affinity Purification (iLAP) plates are an innovative, patent-pending product that combines the cell lysis and HIS-tagged protein purification steps into one. The high-density coating on the plate includes both the CelLytic™ cell lysis reagent and the High Capacity HIS-Select nickel chelate matrix. The CelLytic component of the plate also includes lysozyme for cell lysis of both gram-positive and gram-negative bacteria, protease inhibitors to help prevent proteolytic degradation of target proteins, and benzonase to clarify viscous DNA fragments. As the cells are lysed, the target protein is captured using the same highly selective technology as the HIS-Select Affinity Gel. The High Capacity feature of the 96 well plates enables capture of at least 4 µg of HIS-tagged protein per well, making these plates ideal for rapid colony screening or protein:protein interaction assays. Two-milliliter overnight cultures can be grown in 96 DeepWell, 2 ml plates (P1492) and easily be transferred to the iLAP plates for lysis, purification and analysis.

• One-step cell lysis and HIS-tagged protein purification
• Efficient cell lysis without harvesting cells from the culture
• HIS-Select — highly selective for HIS-tagged proteins, less non-specific binding, and high binding capacity
• Ideal for rapid colony screening

One-step Cell Lysis and Purification 100 µl E. coli culture expressing histidine-tagged protein was added to each well of the iLAP plate. Plate was incubated at room temperature for 2 hours to lyse the cells and capture the protein. Plate was washed and protein from one random well was eluted with 250 mM imidazole for analysis. Gel was stained with ProteoSilver™ Silver Stain Kit (PROT-SIL1). Lane 1: E. coli culture Lane 2: Eluted protein

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The HIS-Select HC 96-well plate (S5563) is coated with a proprietary high-density nickel chelate matrix. This patent pending high capacity coating allows a greater binding capacity per-well than any other commercial histidine binding plates (Figure 1).

• Highest binding capacity of histidine-tagged proteins available:
  > 4 µg/well in 96-well plates;
  
• In well quanitation of bound protein
• 70% reduction in cost versus magnetic beads
• Pre-treated to reduce non-specific binding
• Suitable for applications including:
  Expression Screening, PAGE, MALDI-MS, and High Throughput Purification

Figure 1:

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HIS-Select HS coated plates are designed for low level detection of recombinant fusion proteins with histidine-containing tags. The captured proteins can be detected using standard enzyme-linked assay (ELISA) techniques.

• High Sensitivity: < 1 ng/well
• Pre-treated to reduce non-specific binding
• Suitable for ELISA and high throughput applications

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The HIS-Select™ Filter Plate is an immobilized metal-ion affinity chromatography (IMAC) product that allows rapid, high throughput purification of small-scale crude cell extracts containing histidine tagged proteins. The HIS-Select Filter Plate has 96 wells, which contain 20 µm, spherical silica particles (100 nm pore size) with a hydrophilic layer. The silica is derivatized with a proprietary, patented, quadridentate chelate charged with nickel. The HIS-Select Filter Plate is selective for recombinant proteins with histidine tags and exhibits very low non-specific binding of other proteins. The binding capacity of the HIS-Select Filter Plate is typically greater than 2 mg per well as determined with an approximately 30 kDa histidine tagged protein.

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