Biolegend, 914209, PerCP/Cyanine5.5 anti-Pan-Cytokeratin, 25tests

AE-1 immunoreacts with an antigenic determinant present on most of the subfamily A cytokeratins, including cytokeratins with molecular weights of 56.5, 50, 48 and 40 kD. AE-3 reacts with an antigenic determinant shared by the subfamily B cytokeratins including cytokeratins with molecular weights of of 64, 59, 58, 56, and 52 kD.
25tests
Verified Reactivity: Human, Rat
Reported Reactivity: Dog, Non-Human Primate
Antibody Type: Monoclonal
Host Species: Mouse
Immunogen: This antibody was developed using human epidermal keratins.
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and BSA (origin USA)
Preparation: The antibody was purified by affinity chromatography and conjugated with PerCP/Cyanine5.5 under optimal conditions.
Concentration: Lot-specific (to obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.)
Storage & Handling: The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application: ICFC - Quality tested SB - Community Verified
Recommended Usage: Each lot of this antibody is quality control tested by intracellular immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is 5 µL per million cells in 100 µL staining volume or 5 µL per 100 µL of whole blood. It is recommended that the reagent be titrated for optimal performance for each application. * PerCP/Cyanine5.5 has a maximum absorption of 482 nm and a maximum emission of 690 nm.
Excitation Laser: Blue Laser (488 nm)
Additional Product Notes: For use of this antibody in intracellular flow cytometry (ICFC), fixation/permeabilization using Cyto-Fast™ Fix/Perm Buffer Set (Cat. No. 426803) is recommended. For the use of this antibody in spatial biology (SB), we have partnered with Bruker Spatial Biology Biosciences for demonstration of this antibody on their next-generation ChipCytometry instrument called the CellScape™. The CellScape platform is an end-to-end solution for highly multiplexed spatial omics. Combining an advanced, purpose-built imaging system with easy-to-use fluidics for walk-away automation, the CellScape system will accelerate your exploration into the rapidly evolving field of spatial biology. More information on the the Bruker Spatial Biology CellScape and a complete list of our antibodies that have been demonstrated on the instrument can be found here.
RRID: AB_3662341 (BioLegend Cat. No. 914209) AB_3662341 (BioLegend Cat. No. 914210)
Biology Area: Cell Biology, Cell Motility/Cytoskeleton/Structure, Neuroscience, Neuroscience Cell Markers
Molecular Family: Intermediate Filaments
Gene ID: NA
UniProt: View information about Pan-Cytokeratin on UniProt.org
Clone: AE-1/AE-3
Regulatory Status: RUO
Other Names: Pan-Cytokeratin (AE-1/AE-3), AE1/AE3
Isotype: Mouse IgG1, κ
Q: How stable is PerCP/Cyanine5.5 tandem as compared to PerCP alone?
A: PerCP/Cyanine5.5 is quite photostable and also better than PerCP alone in withstanding fixation.
Q: If an antibody clone has been previously successfully used in IBEX in one fluorescent format, will other antibody formats work as well?
A: It’s likely that other fluorophore conjugates to the same antibody clone will also be compatible with IBEX using the same sample fixation procedure. Ultimately a directly conjugated antibody’s utility in fluorescent imaging and IBEX may be specific to the sample and microscope being used in the experiment. Some antibody clone conjugates may perform better than others due to performance differences in non-specific binding, fluorophore brightness, and other biochemical properties unique to that conjugate.
Q: Will antibodies my lab is already using for fluorescent or chromogenic IHC work in IBEX?
A: Fundamentally, IBEX as a technique that works much in the same way as single antibody panels or single marker IF/IHC. If you’re already successfully using an antibody clone on a sample of interest, it is likely that clone will have utility in IBEX. It is expected some optimization and testing of different antibody fluorophore conjugates will be required to find a suitable format; however, legacy microscopy techniques like chromogenic IHC on fixed or frozen tissue is an excellent place to start looking for useful antibodies.
Q: Are other fluorophores compatible with IBEX?
A: Over 18 fluorescent formats have been screened for use in IBEX, however, it is likely that other fluorophores are able to be rapidly bleached in IBEX. If a fluorophore format is already suitable for your imaging platform it can be tested for compatibility in IBEX.
Q: The same antibody works in one tissue type but not another. What is happening?
A: Differences in tissue properties may impact both the ability of an antibody to bind its target specifically and impact the ability of a specific fluorophore conjugate to overcome the background fluorescent signal in a given tissue. Secondary stains, as well as testing multiple fluorescent conjugates of the same clone, may help to troubleshoot challenging targets or tissues. Using a reference control tissue may also give confidence in the specificity of your staining.
Q: How can I be sure the staining I’m seeing in my tissue is real?
A: In general, best practices for validating an antibody in traditional chromogenic or fluorescent IHC are applicable to IBEX. Please reference the Nature Methods review on antibody based multiplexed imaging for resources on validating antibodies for IBEX.