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Alexa Fluor® 488 anti-Cytokeratin 18 Antibody

Pricing & Availability
Clone
DA-7 (See other available formats)
Regulatory Status
RUO
Other Names
Keratin 18, Keratin type I cytoskeletal 18
Isotype
Mouse IgG1, κ
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Product Citations
publications
a-DA-7_AF488_antiCytokeratin18_Antibody_121715
MCF-7 cells were fixed with 1% paraformaldehyde (PFA) for ten minutes at 37°C, permeabilized with 0.5% Triton X-100 buffer for ten minutes at room temperature, and blocked for 30 minutes at room temperature with 5% fetal bovine serum (FBS). Then the cells were stained with 5 µg/mL anti-Cytokeratin 18 (clone DA-7) Alexa Fluor® 488 (green). Nuclei were counterstained with DAPI (blue). Images were acquired with a 60X objective.
  • a-DA-7_AF488_antiCytokeratin18_Antibody_121715
    MCF-7 cells were fixed with 1% paraformaldehyde (PFA) for ten minutes at 37°C, permeabilized with 0.5% Triton X-100 buffer for ten minutes at room temperature, and blocked for 30 minutes at room temperature with 5% fetal bovine serum (FBS). Then the cells were stained with 5 µg/mL anti-Cytokeratin 18 (clone DA-7) Alexa Fluor® 488 (green). Nuclei were counterstained with DAPI (blue). Images were acquired with a 60X objective.
  • b-DA-7_AF488_Cytokeratin18_Antibody_2_101024
    Multiplexed IHC staining of Alexa Fluor® 488 anti-CK18 (clone DA-7) on formalin-fixed paraffin-embedded human colon tissue, validated for use on the Cellscape™. The tissue was iteratively stained with Alexa Fluor® 488 anti-CK18 (clone DA-7, green) and PerCP/Cyanine5.5 anti-EpCAM (Cat. No. 324213, red) for one hour at room temperature. Nuclei were counterstained with DAPI. Images were captured with a 20X objective. Scale bar: 50 µm
Compare all formats See Alexa Fluor® 488 spectral data
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628408 100 µg £238
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Description

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.

Product Details
Technical Data Sheet (pdf)

Product Details

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.

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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)
  1. Taylor-Papadimitriou J, et al. 1989. J. Cell Sci. 94:403.
  2. Lauerova L, et al. 1988. Hybridoma 7:495.
Product Citations
  1. Mikolajczyk SD, et al. 2011. J Oncol. 252361:2011. PubMed
RRID
AB_2566631 (BioLegend Cat. No. 628408)

Antigen Details

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 View all products for this Gene ID
UniProt
View information about Cytokeratin 18 on UniProt.org

Related FAQs

If an antibody clone has been previously successfully used in IBEX in one fluorescent format, will other antibody formats work as well?

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.

Will antibodies my lab is already using for fluorescent or chromogenic IHC work in IBEX?

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.

Are other fluorophores compatible with IBEX?

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.

The same antibody works in one tissue type but not another. What is happening?

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.

How can I be sure the staining I’m seeing in my tissue is real?

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.

Go To Top Version: 3    Revision Date: 10/10/2024

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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