TotalSeq™-Bn1360 anti-CD324 (E-Cadherin) Antibody

Pricing & Availability
Clone
4A2 (See other available formats)
Regulatory Status
RUO
Other Names
Cadherin-1, CAM 120/80, Epithelial cadherin, Uvomorulin, CD_antigen, CD324 Antigen, CDHE, ECAD, LCAM
Isotype
Mouse IgG1, κ
Barcode Sequence
GTGCATTTCACTCAC
Ave. Rating
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Product Citations
publications
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866703 10 µg £253
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Description

E-cadherin is a calcium-dependent cell adhesion molecule (CAM) critical for formation and functions of adherent junctions during development and tumorigenesis. E-cadherin is usually expressed and localized at the surface of epithelial cells. However, it is also expressed in the adult and embryonic forebrain germinal zones in vivo and in cells derived from these regions growing in vitro. Conditional loss of E-cadherin in mouse brain causes defects in the self-renewal of neural stem cells both in vivo and in vitro. Overexpression of E-cadherin increases the number of neural stem cell colonies indicating that E-cadherin is required for the neural stem cell self-renewal function. In human glioblastoma tissue, the expression of E-cadherin is increased and usually correlates with unfavorable outcome.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human, Rat
Antibody Type
Monoclonal
Host Species
Mouse
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and EDTA
Preparation
The antibody was purified by chromatography and conjugated with TotalSeq™-Bn oligomer under optimal conditions.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C. Do not freeze.
Application

SB - Quality tested

Recommended Usage

Each lot of this antibody is quality control tested by immunofluorescent staining in formalin-fixed, paraffin-embedded (FFPE) lymphoid tissue, and the oligomer sequence is confirmed by sequencing. TotalSeq™-Bn antibodies are compatible with the 10x Visium CytAssist Gene and Protein Expression Assay.

To maximize performance, it is strongly recommended that the reagent be titrated for each application, and that you centrifuge the antibody dilution at 14,000xg at 2 − 8°C for 10 minutes before use. Carefully pipette out the liquid avoiding the bottom of the tube when handling. To determine and optimize dilutions for the addition of Totalseq™-Bn antibodies into pre-designed antibody panels, refer to 10x Genomics Custom Add-on Antibody Optimization guide.


Buyer is solely responsible for determining whether Buyer has all intellectual property rights that are necessary for Buyer's intended uses of the BioLegend TotalSeq™ products. For example, for any technology platform Buyer uses with TotalSeq™, it is Buyer's sole responsibility to determine whether it has all necessary third party intellectual property rights to use that platform and TotalSeq™ with that platform.
Application Notes

Lower MW bands detected in WB are cleaved C terminal fragments.

Cleavage of E-cadherin usually occurs in tumor development and cleaved E-cadherin fragments of ~29-38 kD often possess distinct oncogenic properties.

Additional Product Notes

TotalSeq™-Bn reagents are designed to profile protein levels following an optimized protocol in spatial transcriptomics. Compatible spatial biology devices (e.g. Imaging System, 10x Genomics Visium Spatial CytAssist Gene and Protein Expression instruments and reagents) and sequencer (e.g. Illumina analyzers) are required. TotalSeq™-B reagents are not compatible with the 10x Genomics Visium system. The complete barcode sequence may be provided upon request. Please contact technical support for more information, or visit TotalSeq™-Bn Reagents for 10x Genomics Visium CytAssist Gene and Protein Assay.

Application References

(PubMed link indicates BioLegend citation)
  1. David, J.M. and Rajasekaran, A.K. 2012. Cancer Res. 72(12): 2917–2923.
RRID
AB_3097338 (BioLegend Cat. No. 866703)

Antigen Details

Structure
Human CD324 (E-cadherin) is a 882 amino acid protein with a predicted molecular mass of 97 kD and observed molecular mass of ~120 kD.
Distribution

Tissue distribution: Non-neural epithelial tissues and forebrain germinal zones
Cellular distribution: Golgi, plasma membrane, endosome, and cell

Function
CD324 (E-cadherin) is involved in cell adhesion.
Interaction
Interacts with catenin family members
Cell Type
Embryonic Stem Cells, Epithelial cells, Neural Stem Cells
Biology Area
Cell Adhesion, Cell Biology, Immuno-Oncology, Immunology, Neuroscience, Stem Cells
Molecular Family
Adhesion Molecules, CD Molecules
Antigen References
  1. Lewis-Tuffin LJ, et al. 2010. PLoS One. 5(10):e13665.
  2. Karpowicz P, et al. 2009. J Neurosci. 29(12):3885-96.
Gene ID
999 View all products for this Gene ID
UniProt
View information about CD324 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: 1    Revision Date: 11/17/2023

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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