FACS Antibodies
Fluorescence-activated cell sorting (FACS) is a specialized type of flow cytometry. It sorts a heterogeneous mixture of biological cells into two or more containers, one cell at a time, based upon the specific light scattering and fluorescent characteristics of each cell and its attached antibodies.
- How to choose an antibody for FACS experiment?
- Fluorescent Conjugate Antibodies for FACS Sorted after emission
- Cell Surface Marker Eg: CD3, CD45, CD56
- FACS Antibodies for Immuno Oncology Eg: PD-1, CD30
- Additional Reading
Find your conjugated FACS Antibodies! Choose "all filter options" then your desired conjugate
How to choose Antibody for FACS Experiment?
Follow this guide to accurate flow cytometry results. In a flow cytometry experiment antibodies play a crucial role. Antibodies conjugated to fluorescent markers are able to identify our specific cells of interest and quantitate the amount of our target on the cell.
There are several different production methods for antibodies, each with their positives and negatives. The antibodies differ in binding quality and ultimately the staining of the cells. Polyclonal antibodies are more likely to cross-react with proteins other than the target and are susceptible to variation between batches. Monoclonals, especially recombinant ones, are consistent and reproducible. Specifically, monoclonal antibodies bind to the same epitope, making them ideal for flow cytometry experiments. They reduce the chance of off-target binding and delivering the highest possible batch-to-batch reproducibility.
Recombinant antibodies are a serious alternative as well. Since they are genetically engineered, it is possible to change the Fc portion of the antibody, reducing or even eliminating binding to the Fc Receptors on target cells. Caution, not all desired targets are currently covered by recombinant antibodies.
When we narrow down our pool of possible antibodies to monoclonals we still have to distinguish between different clone lines. Not all are suited for flow cytometry and some even might recognize the wrong epitope, such as a product targeting an intracellular epitope when the intention is to avoid permeabilizing the cells.
One last question that remains is whether to use a directly conjugated or unconjugated antibody. Direct conjugates simplify the assay protocol, reducing the potential for mistakes to occur during sample preparation. Especially for multiplexing approaches they are the more suitable format - allowing simultaneous use of multiple antibodies raised in the same species and enabling increased experimental complexity. If the desired antibody-fluorophore combination is unavailable though, primary antibodies combined with conjugated secondary antibodies can be utilized as substitute.
Additional Reading
- Fluorescent Conjugate Antibodies Sorted after emission peak
- Flow Cytometry Guide Principle of FACS analysis
- Introduction to Fluorescence Focus on life-sciences usage
- Five Tips for choosing the perfect Antibody For general experimental design
Fluorescent Conjugate Antibodies for FACS
Fluophore | Excitation Peak (nm) | Emission Peak (nm) | Emission Color |
400 | 421 | ![]() |
|
401 | 421 | ||
352 | 435 | ||
346 | 442 | 390 | 470 |
493 | 519 | ||
492 | 520 | ||
501 | 523 | ||
550 | 570 | ||
550 | 570 | ||
Phycoerythrin (PE) | 565 | 578 | |
563 | 592 | ||
591 | 616 | ||
629 | 657 | ||
APC | 650 | 661 | |
651 | 667 | ||
650 | 670 | ||
PerCp | 490 | 675 | |
633 | 684 | ||
Cy5.5 | 675 | 694 | |
684 | 702 | ||
680 | 704 | ||
778 | 754 | ||
771 | 796 | ||
784 | 814 |
Cell Surface Marker
Sell-surface markers automatically come to mind in the search for FACS antibodies. The antibodies are usually directed against specific surface proteins (e.g. proteins of the CD classification; CD = cluster of differentiation). After labeling, sorting can then also take place according to these characteristics.
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FACS Antibodies for Immuno Oncology
Immunophenotyping is a technique used to study the protein expressed by cells. An example is the detection of tumor marker, such as in the diagnosis of leukemia. It involves the labelling of white blood cells with antibodies directed against surface proteins on their membrane. By choosing appropriate antibodies eg CD19, CD10, CD45 or HLA-DR the differentiation of leukemic cells can be accurately determined. The labelled cells are then processed in a flow cytometer. The whole procedure can be performed on cells from the blood, bone marrow or spinal fluid.
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Can't find the right antibody? If you need help to find a suiting antibody for flow cytometry, please contact our customer support via phone, live chat or email.