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Assay Cascade Protocols
Frederick National Lab
Nanotechnology Characterization Laboratory

Physicochemical Characterization Method ID

Size, Size Distribution

    Measuring the Size of Nanoparticles in Aqueous Media Using Batch-Mode DLS
Old Television Clip Art See the DLS Video Protocol demonstration
    Size Measurement of Nanoparticles Using Atomic Force Microscopy NIST-NCL PCC-6
    Measuring the Size of Nanoparticles Using Transmission Electron Microscopy NIST-NCL PCC-7
    Analysis of Gold Nanoparticles by Electrospray Differential Mobility Analysis NIST-NCL PCC-10
    Measuring the Size of Colloidal Gold Nanoparticles Using High-Resolution Scanning Electron Microscopy NIST-NCL PCC-15
Molecular Weight
Chemical Composition
    Determination of Gold in Rat Tissue with Inductively Coupled Plasma-Mass Spectrometry NIST-NCL PCC-8
    Determination of Gold in Rat Blood with Inductively Coupled Plasma-Mass Spectrometry NIST-NCL PCC-9
    Method for Determination of the Mass Fraction of Particle-Bound Gold in Suspensions of Gold Nanoparticles NIST-NCL PCC-11
    Quantification of Free and Chelated Gadolinium Species in Nanoemulsion-Based Magnetic Resonance Imaging Contrast Agent Formulations using Hyphenated Chromatography Methods NIST-NCL PCC-14
  Surface Characteristics
Zeta Potential
    Measuring Zeta Potential of Nanoparticles PCC-2
    Measuring the Electrolytic Conductivity of Nanoparticle Suspensions NIST-NCL PCC-12
    Measuring the pH of Nanoparticle Suspensions NIST-NCL PCC-13

In Vitro Characterization Method ID
    Detection of Endotoxin Contamination  
      End Point Chromogenic LAL Assay STE-1.1
      Kinetic Turbidity LAL Assay STE-1.2
      Gel-Clot LAL Assay STE-1.3
      Kinetic Chromogenic LAL Assay STE-1.4
    Detection of Microbial Contamination STE-2.1
    Detection of Bacterial Contamination STE-2.2
    Detection of Mycoplasma Contamination STE-3
    Cell Binding/Internalization  
  Drug Release  
    In Vitro Blood Partitioning Assay GTA-13
  In Vitro Immunology  
    Blood Contact Properties  
      Analysis of Hemolytic Properties of Nanoparticles ITA-1
      Analysis of Platelet Aggregation ITA-2
      Analysis of Nanoparticle Interaction with Plasma Proteins by 2D PAGE ITA-4
      Qualitative Analysis of Total Complement Activation by Western Blot ITA-5.1
      Quantitative Analysis of Complement Activation ITA-5.2
      Coagulation Assay ITA-12
    Cell-Based Assays  
      Mouse Granulocyte Macrophage Colony-Forming Unit Assay ITA-3
      Leukocyte Proliferation Assay ITA-6
      Macrophage/Neutrophil Function (4 categories):  
        Detection of Nitric Oxide Production by RAW 264.7 Macrophage Cell Line ITA-7
        Chemotaxis Assay ITA-8
        Phagocytosis Assay ITA-9
      Measurement of Nanoparticle Effects on Cytotoxic Activity of NK Cells by Label-Free RT-CES System ITA-11
      Analysis of Nanoparticle Effects on Maturation of Monocyte Derived Dendritic Cells In Vitro ITA-14
      In Vitro Induction of Leukocyte Procoagulant Activity by Nanoparticles ITA-17
    Oxidative Stress  
      Hep G2 Hepatocyte Glutathione Assay GTA-3
      Hep G2 Hepatocyte Lipid Peroxidation Assay GTA-4
      Hepatocyte Primary ROS Assay GTA-7
    Cytotoxicity (necrosis)  
      LLC-PK1 Kidney Cytotoxicity Assay (MTT and LDH Release) GTA-1
      Hep G2 Hepatocarcinoma Cytotoxicity Assay (MTT and LDH Release) GTA-2
    Cytotoxicity (apoptosis)  
      LLC-PK1 Kidney Apoptosis Assay (Caspase 3 Activation) GTA-5
      Hep G2 Hepatocarcinoma Apoptosis Assay (Caspase 3 Activation) GTA-6
      Hep G2 Hepatocarcinoma Homogeneous Apoptosis Assay (Caspase 3/7 Activation) GTA-14
      Autophagic Dysfunction Assay: Qualitative Analysis of MAP LC3I to LC3-II Conversion by Western Blot GTA-11
      Autophagic Dysfunction in LLC-PK1 Cells GTA-12

In Vivo Characterization

  Disposition study  
    Tissue Distribution  
    Systemic exposure (plasma AUC)  
  Single and Repeat-Dose Toxicity  
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