Kitos Biotech srls

To determine protein levels of your target of interest. We perform protein extraction and western blot with commercial antibodies to compare protein levels in distinct cell lines or to evaluate changes in protein expression following treatment with your compound of interest. Cell lines can be chosen from our in-house panel or shipped from you to our laboratory.

Features of the assay

• We support you designing the experimental plan. Based on more than 10 years of experience and more than 500 western blot performed, we can help you deciding which antibody to order and test, which cell lines to include as control, and which protocol best fits your target.
  
• We perform the entire experimental plan and analysis in our laboratory, from compound treatment of cells to image acquisition.
  
• We support you with data interpretation.

We offer a robust assay to determine kinetic of compound potency and efficacy based on live-cell imaging.

Features of the assay

• Number of live/dead cells can be measured multiple times over a short (time-lapse) or a long period of treatment.
  
• IC50, relative growth and percentage of growth inhibition (IC50, GI50, TGI and LC50) calculations.
  
• Available in high-throughput (HTS).
  
• Accurate
  
  • Number of cells is estimated by counting nuclei.
    
  • Fraction of dead cells is excluded by live cell counting.
    
  • Multiple reads of the same plate allow well-to-well normalized data analysis.
    
• Informative on the mechanism of action
  
  • It discriminates between cytostatic and cytotoxic compounds.
    
  • Phase contrast images can be available on request to evaluate cell morphology.

We offer a robust assay to determine kinetic of compound potency and efficacy based on live-cell imaging.

Features of the assay

• Number of live/dead cells can be measured multiple times over a short (time-lapse) or a long period of treatment.
  
• IC50, relative growth and percentage of growth inhibition (IC50, GI50, TGI and LC50) calculations.
  
• Available in high-throughput (HTS).
  
• Accurate
  
  • Number of cells is estimated by counting nuclei.
    
  • Fraction of dead cells is excluded by live cell counting.
    
  • Multiple reads of the same plate allow well-to-well normalized data analysis.
    
• Informative on the mechanism of action
  
  • It discriminates between cytostatic and cytotoxic compounds.
    
  • Phase contrast images can be available on request to evaluate cell morphology.

We offer a citotoxicity and proliferation assay to determine kinetic of compound efficacy on 3D tumor spheroids based on live-cell imaging.

Features of the assay

• Spheroid size and cell death can be measured multiple times over a short (time-lapse) or a long period of time.
  
• Available in high-throughput (HTS).
  
• Accurate
  
  • A single spheroid forms in each well of the plate. Spheroid sizes are estimated by measuring their area and calculating the volume.
    
  • Multiple reads at various time points are carried out to obtain spheroid area values normalized on initial spheroid sizes.
    
• Informative on the mechanism of action
  
  • It discriminates between cytostatic and cytotoxic compounds.

An alternative method to flow cytometry to assess cell cycle using fluorescence microscopy and cell imaging.

Features of the assay

• Available for high-content analysis.
  
• Quan­titative, fully automated and unbiased cell cycle distributions.
  
• It can be multiplexed with other of our cell-based assays.

We offer a citotoxicity and proliferation assay to determine kinetic of compound efficacy on 3D tumor spheroids based on live-cell imaging.

Features of the assay

• Spheroid size and cell death can be measured multiple times over a short (time-lapse) or a long period of time.
  
• Available in high-throughput (HTS).
  
• Accurate
  
  • A single spheroid forms in each well of the plate. Spheroid sizes are estimated by measuring their area and calculating the volume.
    
  • Multiple reads at various time points are carried out to obtain spheroid area values normalized on initial spheroid sizes.
    
• Informative on the mechanism of action
  
  • It discriminates between cytostatic and cytotoxic compounds.

We offer a citotoxicity and proliferation assay to determine kinetic of compound efficacy on 3D tumor spheroids based on live-cell imaging.

Features of the assay

• Spheroid size and cell death can be measured multiple times over a short (time-lapse) or a long period of time.
  
• Available in high-throughput (HTS).
  
• Accurate
  
  • A single spheroid forms in each well of the plate. Spheroid sizes are estimated by measuring their area and calculating the volume.
    
  • Multiple reads at various time points are carried out to obtain spheroid area values normalized on initial spheroid sizes.
    
• Informative on the mechanism of action
  
  • It discriminates between cytostatic and cytotoxic compounds.

We offer a reliable and broadly employed cell viability assay to determine compound efficacy.

Features of the assay

• IC50 calculation.
  
• Number of viable cells in culture is estimated with Promega Cell-Titer Glo® assay.

We offer a cell-based assay to determine compound-mediated induction of apoptosis by measuring relative enzymatic activities of caspase 3/7, 8 and 9 in cell culture.

Features of the assay

• Caspases activities are independently measured with Promega Caspase-Glo® 3/7, 8 or 9 assays.
  
• Induction of apoptosis can be correlated with cell viability by multiplexing the assay with our kinetic of cytotoxicity and proliferation assay.

We offer a cell-based assay to measure anti-oxidant activity of tested compound by measuring relative levels of hydrogen peroxide, a reactive oxigen species, in cell culture.

Features of the assay

• Levels of H2O2 are measured with Promega ROS-Glo® H2O2 assay.
  
• Antioxidant activity can be correlated with cell viability by multiplexing the assay with our kinetic of cytotoxicity and proliferation assay.

We offer a cell-based assay to determine potency of histone deacetylase (HDAC) class I and II inhibitors by measuring relative enzymatic activity of HDAC I/II in cell culture.

Features of the assay

• Activities of HDAC I/II are measured with Promega HDAC-Glo® assay.
  
• HDAC inhibitor potency can be correlated with cell viability by multiplexing the assay with our kinetic of cytotoxicity and proliferation assay.

We offer a citotoxicity and proliferation assay to determine contribution of ABC transporters to cancer cell chemoresistance of tested compound based on live-cell imaging.

Features of the assay

• Cells are treated with three selective inhibitors of ABC transporters together with tested compound.
  
• Number of live/dead cells is measured with our kinetic of cytotoxicity and proliferation assay.
  
• Relative cell number, relative growth and percentage of growth inhibition calculations.

We offer a cell-based screening to test compound libraries of your choice in your model of interest.

Features of the assay

• We have available various commercial libraries of known compounds from Selleckchem:

  • PI3K/Akt Inhibitor Library - 118 inhibitors
    
  • Epigenetics Compound Library - 182 small molecules
    
  • Anti-cancer Compound Library - 422 compounds
    
  • Apoptosis Compound Library - 101 small molecules
    
  • FDA-approved Drug Library - 1443 drugs
    
  • Tyrosine Kinase Inhibitor Library - 171 inhibitors
    
  • Autophagy Compound Library - 154 inhibitors
    
  • MAPK Inhibitor Librabry - 61 inhibitors
    
  • Natural Product Library - 133 products
    
  • Stem Cell Signaling Compound Library - 89 inhibitors
    
  • Protease Inhibitor Library - 53 inhibitors
    

• We can test these libraries in our cell lines or in cells provided by customers.

• We can employ any of our assays available in high-throughput:

  • Kinetic of Cytotoxicity and Proliferation assay
    
  • 3D Spheroid Drug Sensitivity assay
    
  • Cell Viability assay
    
  • Antioxidant assay
    
  • Histone Deacetylase Activity assay
    

• We can evaluate the combinatorial efficacy of your favorite compound with these libraries to perform drug combination studies.

• Selected hits will be validated with the corresponding assay with 11 concentrations of compound.

Protein Services

Protein Expression Visualization Services