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Purified 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
Ave. Rating
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Product Citations
publications
A_4A2_PURE_CD324_Antibody_082719
Western blot of purified anti-CD324 (E-Cadherin) antibody (clone 4A2). Lane 1: Molecular weight marker; Lane 2: 30 µg of human skin lysate; Lane 3: 30 µg of rat colon lysate. The blot was incubated with 5 µg/mL of the primary antibody overnight at 4°C, followed by incubation with HRP-labeled goat anti-mouse IgG (Cat. No. 405306). Enhanced chemiluminescence was used as the detection system. The top arrow indicates the full length E-Cadherin protein. The bottom arrow indicates the 33-kDa C-terminal fragment (CTF2).
  • A_4A2_PURE_CD324_Antibody_082719
    Western blot of purified anti-CD324 (E-Cadherin) antibody (clone 4A2). Lane 1: Molecular weight marker; Lane 2: 30 µg of human skin lysate; Lane 3: 30 µg of rat colon lysate. The blot was incubated with 5 µg/mL of the primary antibody overnight at 4°C, followed by incubation with HRP-labeled goat anti-mouse IgG (Cat. No. 405306). Enhanced chemiluminescence was used as the detection system. The top arrow indicates the full length E-Cadherin protein. The bottom arrow indicates the 33-kDa C-terminal fragment (CTF2).
  • B_4A2_PURE_CD324_Antibody_SB_111323
    SeqIF™ (sequential immunofluorescence) staining on COMET™ of Purified anti-CD324 (E-Cadherin) (clone 4A2, yellow) on formalin-fixed paraffin-embedded human prostate carcinoma tissue at 5 µg/mL. Alexa Fluor™ Plus 647 Goat anti-Mouse IgG antibody (Lunaphore, Cat. No. DR647MS) was used as a secondary antibody. Nuclei were counterstained with DAPI (blue). Tissue underwent an all-in-one dewaxing and antigen retrieval preprocessing.
  • C_4A2_PURE_CD324_Antibody_3_031324
    IHC staining of Purified anti-CD324 (E-Cadherin) (clone 4A2) on formalin-fixed paraffin-embedded human tonsil tissue. Following antigen retrieval using Sodium Citrate H.I.E.R., 1X (Cat. No. 928502), the tissue was incubated without (panel A) and with (panel B) 10 µg/mL of Purified anti-CD324 (E-Cadherin) (clone 4A2) followed by incubation with Alexa Fluor® 647 Goat anti-mouse IgG (Cat. No. 405322) for 1 hour at room temperature. Nuclei were counterstained with DAPI (Cat. No. 422801). Images were captured with a 40X objective. Scale bar: 50 µm
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866701 25 µg $106
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866702 100 µg $276
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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

Product Details

Verified Reactivity
Human, Rat
Antibody Type
Monoclonal
Host Species
Mouse
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
IHC-P - Verified
SB - Community verified
Recommended Usage

Each lot of this antibody is quality control tested by Western blotting. For Western blotting, the suggested use of this reagent is 1.0 - 10 µg per mL. For immunohistochemistry on formalin-fixed paraffin-embedded tissue sections, a concentration range of 1 - 10 µg/mL is suggested. It is recommended that the reagent be titrated for optimal performance for each application.

It is also recommended using EDTA-based solutions for dissociating attachment-dependent cell lines.

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

For the use of this antibody in spatial biology applications, we have partnered with Lunaphore Technologies for demonstration of our antibodies on the COMET™. The COMET™ platform is an automated, end-to-end spatial biology solution developed for rapid and flexible multiplex tissue profiling. More information on the COMET™ and a complete list of our antibodies that have been demonstrated on the COMET™ can be found here.

Application References

(PubMed link indicates BioLegend citation)
  1. David, J.M. and Rajasekaran, A.K. 2012. Cancer Res. 72(12): 2917–2923.
Product Citations
  1. Lin CY, et al. 2022. Int J Mol Sci. 23:. PubMed
RRID
AB_2814610 (BioLegend Cat. No. 866701)
AB_2814610 (BioLegend Cat. No. 866702)

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
Specificity (DOES NOT SHOW ON TDS):
CD324
Specificity Alt (DOES NOT SHOW ON TDS):
CD324
App Abbreviation (DOES NOT SHOW ON TDS):
WB,IHC-P,SB
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: 3    Revision Date: 03/13/2024

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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