Biolegend, 628408, Alexa Fluor® 488 anti-Cytokeratin 18 Antibody, 100μg

Cytokeratin 18, also known as keratin 18, is a type I intermediate filament protein with a molecular weight of approximately 48 kD. Cytokeratin 18 is a heterotetramer composed of two type I and two type II keratins. Cytokeratin 18 associates with cytokeratin 8 and is widely expressed in simple epithelial tissues in adults, where it is found in the cytoplasm, nucleus, and nucleolus. Cytokeratin 18 is associated with the cytoskeleton and mutations in this protein have been associated with liver disease after environmental insult. Cytokeratin 18 can be modified by glycosylation, phosphorylation (Ser34 and Ser53), acetylation (Ser2), and proteolytic cleavage at amino acids 238 and 397 by caspase family members. The cytokeratin 18 protein interacts with a number of proteins including caspase 3, 14-3-3 gamma, 14-3-3 zeta, 14-3-3 sigma 14-3-3 beta, 14-3-3 eta, keratin 5, keratin 8, TNF receptor II, plakophilin 2, EGF receptor, and usherin, among others.
100μg
Verified Reactivity: Human
Antibody Type: Monoclonal
Host Species: Mouse
Immunogen: Human breast carcinoma cell line PMC-42
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation: The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 488 under optimal conditions.
Concentration: 0.5 mg/ml
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: ICC - Quality tested SB - Community Verified
Recommended Usage: Each lot of this antibody is quality control tested by immunocytochemistry. For immunocytochemistry, a concentration range of 0.1 - 10 μg/ml is recommended. It is recommended that the reagent be titrated for optimal performance for each application. * Alexa Fluor® 488 has a maximum emission of 519 nm when it is excited at 488 nm. Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.View full statement regarding label licenses
Excitation Laser: Blue Laser (488 nm)
Application Notes: The DA-7 monoclonal antibody recognizes human cytokeratin 18 and is useful for Western blotting. Additional reported applications (for the relevant formats) include: immunoprecipitation, immunohistochemistry of paraffin-embedded sections, and ELISA. This clone and BioLegend clone LDK18 displayed similar affinities when tested for western blot applications
Additional Product Notes: 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.
Application References(PubMed link indicates BioLegend citation): Taylor-Papadimitriou J, et al. 1989. J. Cell Sci. 94:403. Lauerova L, et al. 1988. Hybridoma 7:495.
Product Citations: Mikolajczyk SD, et al. 2011. J Oncol. 252361:2011. PubMed
RRID: AB_2566631 (BioLegend Cat. No. 628408)
Structure: Type I intermediate filament protein, contains three coiled-coil domains, approximately 48 kD. Heterotetramer composed of two type I and two type II keratins. Cytokeratin 18 associates with cytokeratin 8.
Distribution: Widely expressed in simple epithelial tissues in adults. Can be found in the cytoplasm, nucleus, and nucleolus
Function: Intermediate filament protein involved with the cytoskeleton. Mutations in cytokeratin 18 may predispose to liver disease upon exposure to some viruses or hepatoxins
Interaction: Interacts with a number of proteins including caspase3, 14-3-3 gamma, 14-3-3 zeta, 14-3-3 sigma 14-3-3 beta, 14-3-3 eta, keratin 5, keratin 8, TNF receptor II, plakophilin 2, EGF receptor, usherin
Modification: Phosphorylation (Ser34, Ser53), glycosylation, acetylation (Ser2). Can also be cleaved at amino acid 238 and 397 by caspase family members
Biology Area: Cell Biology, Cell Motility/Cytoskeleton/Structure, Neuroscience, Neuroscience Cell Markers
Molecular Family: Intermediate Filaments
Antigen References: 1. Ku N, et al. 2003. Proc. Natl. Acad. Sci. 100:6063. 2. Kulesh DA and Oshima RG. 1989. Genomics 4:339. 3. Steinart PM and Roop DR. 1988. Ann. Rev. Biochem. 57:593.
Gene ID: 3875
UniProt: View information about Cytokeratin 18 on UniProt.org
Clone: DA-7
Regulatory Status: RUO
Other Names: Keratin 18, Keratin type I cytoskeletal 18
Isotype: Mouse IgG1, κ
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.