Sample Preparation

Sample preparation is the most important step in any analytical technique for achieving optimal and reproducible results. Orochem offers a wide range of products for efficient sample preparation for down-stream applications like SDS-PAGE, 2-D PAGE, NMR, enzyme assays, mass spectrometry etc. for protein research.

  • Coral Desalting 96-well Spin Plates
  • Coral Sephadex Packed Plates
  • Reva Detergent Removal
  • Reva Lipids Removal
  • Reva Endotoxin Removal
  • Reva Abundant Protein removal
  • Catalog#
  • Description
  • Reva Lipids removal

    Reva Lipids removal (1)

    Reva Lipids Removal

    Orochem’s Reva lipids removal resin is extremely effective for removal of lipids from biological samples such as serum, tissue extracts, protein samples, etc.
    Catalog No. Description Qty/Pk
    BKLR-10 Reva Lipids Removal Resin, 10 ml 10 ml
  • Reva Detergent

    Reva Detergent (1)

    Reva Detergent Removal Resin

    Detergents or surfactants are used extensively in protein chemistry to solubilize and stabilize proteins and to disaggregate protein complexes [1]. However, the presence of excess unbound detergent interferes with many downstream applications like ELISA, enzyme assays, isoelectric focusing, and mass spectrometry. Orochem’s REVA detergent removal resin specifically binds and removes high concentrations of a wide variety of detergents used in protein research for the preparation of biological samples. The resin is designed for the efficient removal of a wide variety of commonly used ionic, nonionic, and zwitterionic detergents from proteins and peptide samples with high sample recovery in a centrifuge format for 25 to 1000 μl samples. Detergent removal is based on the presence of a small hydrophobic cavity in the resin into which appropriately sized non-polar moieties such as detergents can enter to form an inclusion complex. The easy-to-use spin format is fast (less than 15 min) and significantly improves results over the standard drip column and batch methodologies with >95% removal of detergents. The resin is available in bulk resin slurries, 96-well plate format for high-throughput applications as well as in spin column format.

    Highlights

    • Efficient Detergent Removal - removes >95% of detergents
    • Easy-to-use - no cumbersome column preparation or equilibration
    • Fast - no waiting for protein to emerge by gravity-flow
    • High protein recovery - low non-specific protein binding resin maximizes protein recovery
    Reference: [1] Neugebauer, J. (1990). Methods Enzymol. 182, 239-282.
  • Coral Sephadex Packed Plate

    Coral Sephadex Packed Plate (1)

    Coral Sephadex Packed Plates

    Dye - Terminator removal is one of the major cleanup objective to yield a highly purified product prior to conducting a sequencing reaction. OrochemSephadex packed plates provide a very cost effective solution for high performance parallel processing of 96 sequencing reactions by gel filtration.
    • OrochemSephadex packed plates are available in 96-wells and 384-wells formats.
    • These plates can be used with gel filtration media for high throughput sequencing reaction cleanup.
    • They are constructed from rigid polystyrene that can withstand centrifugation.

    General Protocol

    • Place the Sephadex plate over appropriate wash plates and place it into a centrifuge with correct carrier.
    • Centrifuge for 1 minute at 750 ×g to remove residual buffer. Discard the wash plates containing excess water.
    • Carefully and rapidly load 50 µl sequencing reaction mixture before the Spheadex gel dries out.
    • Centrifuge for 2 minutes at 750 ×g to eliminate big dye terminators, salts, and low molecular weight species.
    • Purified DNA of the sequencing reaction is recovered in collection plate
  • Coral Desalting Plate

    Coral Desalting Plate (4)

    Coral Desalting

    Desalting is a critical step in processing samples that have been isolated in the presence of salts and other small molecules. Resin-based desalting is performed by allowing the small molecules to freely enter the resin pore and be retarded in their flow through the packed resin bed while the high molecular weight molecules like proteins in the sample are excluded from the resin and rapidly exit the column. The spin column method eliminates cumbersome column preparation or equilibration, allowing multiple-sample processing in 7,000 Da.

    Features:

    Easy-to-use- no cumbersome column preparation or equilibration Fast- no waiting for protein to emerge by gravity-flow High protein recovery- low non-specific protein binding resin maximizes protein recovery

    Protocol for Desalting or Removing Small Molecules

    • Equilibrate Coral 96-well Desalt Spin Plates to room temperature
    • Remove the sealing material from the bottom of the plate and place it on top of a wash plate.
    • Remove the sealing material from the top of the desalt plate.
    • Place the assembly into a centrifuge with 96-well plate-carrier rotor and centrifuged at 1,000×g for 2 minutes to remove the storage buffer. Discard the flow-through.
    • Rinse the wash plate three times with deionized water, dry and save for future use.
    • Stack the desalt plate on top of a sample collection plate (blue), aligning the alphanumeric indices on the plates.
    • Apply sample (20-100µl) to the center of the resin bed. To expel the entire sample, carefully touch pipette tip to the resin. For 20 µl protein samples (>300 µg/ml), apply a 20 µl stacker of water or buffer on top of resin bed after the sample wash has fully absorbed to ensure maximal protein recovery.
    • Centrifuge the plate assembly at 1,000×g for 2 minutes to collect the desalt sample. Discard the desalt plate or reserve it for further balancing purpose.
Title Citation Cutomer Affiliation
Automated tryptic digestion procedure for HPLC/MS/MS peptide mapping of immunoglobulin gamma antibodies in pharmaceutics Chelius, Dirk, Gang Xiao, Andrew C. Nichols, Alona Vizel, Bing He, Thomas M. Dillon, Douglas S. Rehder et al. Journal of pharmaceutical and biomedical analysis 47, no. 2 (2008): 285-294. Amgen, CA
Immunoglobulin G (IgG) Fab glycosylation analysis using a new mass spectrometric high-throughput profiling method reveals pregnancy-associated changes Bondt, Albert, Yoann Rombouts, Maurice HJ Selman, Paul J. Hensbergen, Karli R. Reiding, Johanna MW Hazes, Radboud JEM Dolhain, and Manfred Wuhrer. Molecular & Cellular Proteomics 13, no. 11 (2014): 3029-3039. Erasmus University Medical center, UV University Amsterdam, The Netherlands,