Proteomics

Orochem Technologies offers a wide range of products for protein research which meets customer’s varied needs of sample preparation, affinity protein purification to protein detection and quantitation. Some of the products offered by Orochem to support your needs for protein research are:

  • High-quality affinity purification resins and kits for purification of antibodies, fusion-tagged proteins and biotin-labeled molecules.
  • Activated supports for immobilization of proteins and other ligands.
  • Resins for the removal of salts and detergents from protein solutions. Orochem’s Coral desalting resin, spin columns and 96-well plates can be used for the efficient removal of salts with excellent protein recovery.
  • State-of-the-art colorimetric assays for quantitation of total protein.
  • A wide variety of total protein stains for visualization of protein bands resolved by gel electrophoresis (SDS-PAGE) including Coomassie based stains and silver stain, the most sensitive colorimetric method for detecting proteins.
  • Protein stains for visualization of protein bands on nitrocellulose and PVDF membranes.
  • Catalog#
  • Description
  • Cell Lysates

    Cell Lysates (1)

    Cell Lysates

    Universal Filter Plate for Bacterial, Insect, and Human Cell Lysates

    An Apparatus for high-throughput filtration of cell lysates, 96-well plate: The clarification of cell requires separation of cell debris, colloids, insoluble precipitates, aggregates, and other contaminants before the target molecule (proteins) is isolated and characterized. Orochem’s filtration plate is a depth microfiltration system that consists of a filter with an open-pore structure to remove cells and cell debris; and a filter–aid material with a tighter pore structure to remove colloidal matter simultaneously to deliver particulate-free feed for down-stream processes.
    "

    Formats

    High-throughput 96-well plate format Formats Small scale single-use depth filtration systems for large sample volumes (up to 2 liters)

    Advantages

    Efficient and reproducible clarification of cell lysates Fast and high-throughput process. Reduce the processing time to 5 min. Good protein recovery Good sample volume recovery Results comparable or superior to high-speed centrifugation

    Procedures:

    A. Additional Materials RequiredVariable-speed bench-top centrifuge with rotor and appropriate carriers capable of handling stacked plates at 1000 x g B. General Procedure for Lysate Filtration Using 96-well plate by Centrifugation

    1. Perform lysate clarification at room temperature or at 4°C.
    2. Place the filter plate on top of a collection plate.
    3. Apply up to 800 µL of cell lysate to each well.
    4. Place the assembly into a centrifuge with appropriate carriers capable of handling stacked plates,balance the centrifuge with appropriate plates, and centrifuge at 1000 x g for 2 minutes to collectthe clarified cell lysate.

    Cell Lysate Filter

    Experimental Data:

    *This work was completed by the Protein Expression Laboratory at the Frederick National Laboratory

    Figure 1.

    SDS Page Results of E. Coli cell lysate by centrifugation and Cell Lysate Filter Plate. For every pair of samples, the left side was clarified using the centrifuge method without Cell Lysate Filter Plate and the right side was clarified using the filtration method. E.Coli growths were harvested by spinning at 4,000 RPM for 20 minutes at 4 °C. Pellets were resuspended in buffer 20/300/1 1:200 PI and lysed using a microfluidizer (LM20), 2 passes each, and the lysate was clarified using the centrifuge method 10,000 x g for 15 minutes and the filtration method 1,000 x g 2 min. Samples (440 µL) were purified using the PhyNexus MEA system. T= Total protein, L=Load protein (soluble), F=Flow through and 1,2,3= Elutions

  • Sample Preparation

    Sample Preparation (13)

    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
  • Filtration

    Filtration (1)

    Proteomics-Filtration

    Do you need a 96-well plate to support affinity resins for nucleotide and antibody extractions? Orochem’s filter plates are designed for oligo and peptide synthesis and extraction of large molecules.

    Features:

    1. Compatible with resins or beads of different particle sizes.
    2. Compatible with different organic solvents.
    3. Support high recovery of nucleotides, peptides and proteins.

    Applications:

    • Support CPG beads for nucleotide extraction.
    • Support IgG and IgA affinity Matrix beads for antibody extraction.
    • Support ion-exchange resins for desalting N- or O- glycans.
    Customers’ new publications using Orochem’s filter plates
    1. Schaffert, Anja, et al. "Minimal B Cell extrinsic IgG glycan modifications of Pro-and anti-inflammatory IgG preparations in vivo." Frontiers in Immunology 10 (2020): 3024.
    2. Plomp, Rosina, et al. "Comparative glycomics of immunoglobulin A and G from saliva and plasma reveals biomarker potential." Frontiers in immunology 9 (2018): 2436.
    3. Vreeker, Gerda CM, et al. "O-and N-glycosylation analysis of cell lines by ultrahigh resolution MALDI-FTICR-MS." International Journal of Mass Spectrometry 448 (2020): 116267.
  • Affinity Purification

    Affinity Purification (36)

    Affinity Purification

    Affinity chromatography is a powerful technique to enrich or purify a protein from other proteins and components in a biological sample like crude cell lysate or other complex samples. The protein of interest is purified by taking advantage of its specific binding properties to an immobilized ligand. Orochem affinity purification products: 1. EZ Link Resin and Kit 2. EZ Pure Protein A Resin and Kit 3. EZ Pure Protein G Resin and Kit 4. EZ Pure Streptavidin Resin and Plate 5. EZ Pure Ni-NTA Resin and Kit 6. EZ Pure Cobalt Resin and Kit 7. EZ Pure Glutathione Resin and Kit
  • Protein Assays

    Protein Assays (15)

    Protein Assays and Quantitation

    Orochem Protein Assays represent the state-of-the-art colorimetric assays for quantitation of total protein. We offer protein assays for a wide variety of applications. Our assays provide exceptional accuracy, compatibility, reproducibility and broad applicability. We also provide stable and sterile filtered bovine serum albumin (BSA) and bovine gamma globulin (BGG) standards.

    Products:

    • BCA Protein Assay Kit
    • Micro BCA Protein Assay Kit
    • Coomassie Protein Assay Kit
    • Orochem PCV Protein Assay Kit
    • BSA Standard
    • BGG Standard
  • Protein Stains

    Protein Stains (4)

    Protein Stains

    Orochem offers a wide variety of total protein stains for in-gel protein detection and visualization. A wide selection of stains developed at Orochem varies in sensitivity, staining time and reversibility.

    Products

    Coomassie Blue Protein Stain Reagent Silver Protein Stain Kit Reversible Protein Stain Kit for Nitrocellulose Membrane Reversible Protein Stain Kit for PVDF Membrane
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,
Comparative performance of four methods for high-throughput glycosylation analysis of immunoglobulin G in genetic and epidemiological research Huffman, Jennifer E., Maja Pučić-Baković, Lucija Klarić, René Hennig, Maurice HJ Selman, Frano Vučković, Mislav Novokmet et al. Molecular & Cellular Proteomics 13, no. 6 (2014): 1598-1610. University of Edinburgh, Edinburgh, UK; Genos Glycoscience Laboratory, Zagreb, Croatia; Magdeburg, Germany; Leiden University Medical Center, Leiden, The Netherlands; University of Split, Split, Croatia; University of Zagreb, Faculty of Pharmacy and Biochemistry, Zagreb, Croatia; Novosibirsk, Russia; Groningen, The Netherlands; VU University Amsterdam, Amsterdam, The Netherlands;
Combining chemical genetics and proteomics to identify protein kinase substrates. Dephoure, Noah, Russell W. Howson, Justin D. Blethrow, Kevan M. Shokat, and Erin K. O'Shea. Proceedings of the National Academy of Sciences of the United States of America 102, no. 50 (2005): 17940-17945. Howard Hughes Medical Institute and Departments of Biochemistry and Biophysics and Cellular and Molecular Pharmacology, University of California, San Francisco, CA