Carrier-free Antibodies for Mass Cytometry
Our scientists are at the bench daily to produce and validate our antibodies, so they have hands-on experience and knowledge of each antibody’s performance.
Cell Signaling Technology (CST) offers carrier-free antibodies to be used in highly multiplexed single-cell analysis and imaging. These antibodies are:
- Validated in relevant applications for specificity, sensitivity and reproducibility
- Free from BSA or other carrier proteins that interfere with heavy metal conjugation
- Available at high concentrations (>1mg/ml) required for conjugation
Selecting Antibodies for CyTOF® Mass Cytometry and MaxPar® Metal Labeling
The ability to analyze multiple markers in techniques such as flow cytometry and fluorescent multiplex immunohistochemistry (IHC) requires the use of antibodies. Spectral overlap limits the number of fluorochromes that can be concurrently used in a cell or tissue fluorescent assay. In addition, tissue autofluorescence and weak signal intensity can make fluorescent IHC assays more challenging. Cycling amplified fluorescent methods (e.g., Tyramide-based methods) can improve signal intensity in tissue imaging, but entail procedures that can potentially damage sensitive epitopes within the tissue sample and require significant optimization.Why are researchers using CyTOF® (time-of-flight mass spectrometry)
Mass cytometry offers advantages for multi-parameter analysis over flow and IHC applications, using metal-tagged antibodies (e.g. antibodies conjugated to rare metals using MaxPar® conjugation kits). By using heavy metals rather than fluorophores, one can take advantage of the low spectral overlap between heavy metal isotopes and simultaneously profile over 30 proteins within a single cell or tissue. Specifically, mass cytometry or CyTOF® uses time-of-flight mass spectrometry to phenotype cells and analyze cell functions, such as signaling. Imaging Mass Cytometry™ combines CyTOF® capabilities with imaging for spatial context. Imaging cytometers, such as the Hyperion™ (from Fluidigm), are able to simultaneously image dozens of biomarkers in FFPE or frozen tissue samples to provide information on tissue architecture, protein co-localization, and cellular interactions.Why validated antibodies are important
When selecting antibodies for metal conjugation, prior validation in relevant applications offers the greatest chance of success. CST antibodies undergo rigorous validation and specificity testing to ensure reproducible, reliable results. For single-cell mass cytometry, such as Helios™ (from Fluidigm), consider selecting from CST's catalog of flow cytometry-validated antibodies. For Imaging Mass Cytometry™, the antibody you choose should be matched to your sample preparation method. For FFPE samples, consider selecting from CST's catalog of IF-paraffin or IHC-validated antibodies. For frozen tissues, consider selecting from our IF-frozen validated antibodies.
Choose from our comprehensive portfolio of high quality antibodies that are:
- Carrier-free formulated ready for metal conjugation
- Available at high concentrations (>1mg/ml)
- Validated in relevant applications such as flow cytometry and IHC
- Gonzalez VD, Samusik N, Chen TJ, Savig ES, Aghaeepour N, Quigley DA, Huang YW, Giangarrà V, Borowsky AD, Hubbard NE, Chen SY, Han G, Ashworth A, Kipps TJ, Berek JS, Nolan GP, Fantl WJ (2018). Commonly Occurring Cell Subsets in High-Grade Serous Ovarian Tumors Identified by Single-Cell Mass Cytometry. Cell Rep. 22(7), 1875-88.
- Schulz D, Zanotelli VRT, Fischer JR, Schapiro D, Engler S, Lun XK, Jackson HW, Bodenmiller B (2018). Simultaneous Multiplexed Imaging of mRNA and Proteins with Subcellular Resolution in Breast Cancer Tissue Samples by Mass Cytometry. Cell Syst. 6(1), 25-36