PerCP/Cyanine5.5 anti-human CD326 (EpCAM) Antibody

Pricing & Availability
Clone
9C4 (See other available formats)
Regulatory Status
RUO
Other Names
Ep-CAM, tumor associated calcium signal transducer 1, epithelial cell surface antigen, epithelial glycoprotein 2, EGP2, adenocarcinoma associated antigen, TROP1
Isotype
Mouse IgG2b, κ
Ave. Rating
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Product Citations
publications
9C4_PerCPCyanine55_CD326_Antibody_110718
Human colon carcinoma cell line HT-29 was stained with CD326 (EpCAM, clone 9C4) PerCP/Cyanine5.5 (filled histogram) or mouse IgG2b, κ PerCP/Cyanine5.5 isotype control (open histogram).
  • 9C4_PerCPCyanine55_CD326_Antibody_110718
    Human colon carcinoma cell line HT-29 was stained with CD326 (EpCAM, clone 9C4) PerCP/Cyanine5.5 (filled histogram) or mouse IgG2b, κ PerCP/Cyanine5.5 isotype control (open histogram).
  • 9C4_PerCPCyanine55_CD326_Antibody_2_101024
    Multiplexed IHC staining of PerCP/Cyanine5.5 anti-CD326 (clone 9C4) on formalin-fixed paraffin-embedded human tonsil tissue, validated for use on the Cellscape™. The tissue was iteratively stained with PerCP/Cyanine5.5 anti-CD326 (clone 9C4, green) and PerCP/Cyanine5.5 anti-CD45 (Cat. No. 368504, red) for one hour at room temperature. Nuclei were counterstained with Hoechst 33342. Images were captured with a 20X objective. Scale bar: 50 µm
Compare all formats See PerCP/Cyanine5.5 spectral data
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324213 25 tests 141€
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324214 100 tests 306€
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Description

CD326 is also known as Ep-CAM, tumor associated calcium signal transducer 1, epithelial cell surface antigen, epithelial glycoprotein 2, EGP2, adenocarcinoma associated antigen, and TROP1. CD326 is a type I transmembrane protein containing six disulfide bridges and one THYRO domain. This cell surface glycosylated 40 kD protein is highly expressed in bone marrow, colon, lung, and most normal epithelial cells and is expressed on carcinomas of gastrointestinal origin. Recently, it has been reported that CD326 expression occurs during the early steps of erythrogenesis. CD326 functions as a homotypic calcium-independent cell adhesion molecule and is believed to be involved in carcinogenesis by its ability to induce genes involved in cellular metabolism and proliferation. CD326 antigen is an immunotherapeutic target for the treatment of human carcinomas.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Human
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
DU.4475 breast carcinoma
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

FC - Quality tested
SB - Community Verified

Recommended Usage

Each lot of this antibody is quality control tested by 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.

* PerCP/Cyanine5.5 has a maximum absorption of 482 nm and a maximum emission of 690 nm.

Excitation Laser
Blue Laser (488 nm)
Application Notes

Additional reported applications (for the revelant formats) include: immunofluorescence, immunohistochemistry3, and spatial biology (IBEX)4,5.

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. Lammers R, et al. 2002. Exp. Hematol. 30:537.
  2. Schultz LD, et al. 2010. P. Natl. Acad. Sci. USA 107:13022. PubMed
  3. Human Protein Atlas http://www.proteinatlas.org/ENSG00000119888/antibody (IHC)
  4. Radtke AJ, et al. 2020. Proc Natl Acad Sci USA. 117:33455-33465. (SB) PubMed
  5. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations
  1. Ge W, et al. 2022. Int J Mol Sci. 24: . PubMed
  2. Lao L, et al. 2023. Cancer Immunol Res. 11:320. PubMed
  3. Otano I, et al. 2021. Nat Commun. 12:7296. PubMed
  4. Cheng L, et al. 2021. Cancer Immunol Immunother. Online ahead of print. PubMed
  5. Gui J, et al. 2011. Int Immunol. 1.407638889. PubMed
  6. Willingham S, et al. 2012. Proc Natl Acad Sci U S A. 109:6627. PubMed
  7. Wang J, et al. 2020. Cell. 183:1264. PubMed
  8. Cai X, et al. 2014. PLoS One. 9:108942. PubMed
  9. Costa A et al. 2018. Cancer cell. 33(3):463-479 . PubMed
  10. Cheah M, et al. 2015. Proc Natl Acad Sci U S A. 112:4725. PubMed
  11. Lau KX, et al. 2020. Nat Commun. 11:2420. PubMed
  12. Uniken Venema WT, et al. 2019. Gastroenterology. 156:812. PubMed
  13. Fridriksdottir A, et al. 2015. Nat Commun. 6: 8786. PubMed
  14. Montel‐Hagen A et al. 2019. Cell stem cell. 24(3):376-389 . PubMed
  15. Kim J, et al. 2018. J Histochem Cytochem. 66:879. PubMed
RRID
AB_2098808 (BioLegend Cat. No. 324213)
AB_2098808 (BioLegend Cat. No. 324214)

Antigen Details

Structure
Type I transmembrane protein, contains six disulfide bridges, one THYRO domain, approximate molecular weight 40 kD.
Distribution

Highly expressed in bone marrow, colon, lung, and most normal epithelial cells. Also highly expressed on carcinomas of gastrointestinal origin. Expressed during early erythrogenesis.

Function
Homotypic calcium-independent cell adhesion. CD326 is believed to be involved in carcinogenesis by its ability to induce genes involved in cellular metabolism and proliferation.
Modification
Glycosylated.
Cell Type
Embryonic Stem Cells, Epithelial cells
Biology Area
Cell Biology, Immunology, Stem Cells
Molecular Family
Adhesion Molecules, CD Molecules
Antigen References

1. Strnad J, et al. 1989. Cancer Res. 49:314.
2. Munz M, et al. 2004. Oncogene 23:5748.
3. Rao CG, et al. 2005. Int. J. Oncol. 27:49.

Gene ID
4072 View all products for this Gene ID
UniProt
View information about CD326 on UniProt.org

Related FAQs

How stable is PerCP/Cyanine5.5 tandem as compared to PerCP alone?

PerCP/Cyanine5.5 is quite photostable and also better than PerCP alone in withstanding fixation.

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.

Other Formats

View All CD326 Reagents Request Custom Conjugation
Description Clone Applications
Purified anti-human CD326 (EpCAM) 9C4 FC,ICC,IHC
FITC anti-human CD326 (EpCAM) 9C4 FC
PE anti-human CD326 (EpCAM) 9C4 FC
APC anti-human CD326 (EpCAM) 9C4 FC
Alexa Fluor® 488 anti-human CD326 (EpCAM) 9C4 FC,SB
Alexa Fluor® 647 anti-human CD326 (EpCAM) 9C4 FC,ICC,SB
PerCP/Cyanine5.5 anti-human CD326 (EpCAM) 9C4 FC,SB
Biotin anti-human CD326 (EpCAM) 9C4 FC
Pacific Blue™ anti-human CD326 (EpCAM) 9C4 FC
Brilliant Violet 421™ anti-human CD326 (EpCAM) 9C4 FC
PE/Cyanine7 anti-human CD326 (EpCAM) 9C4 FC
Brilliant Violet 605™ anti-human CD326 (EpCAM) 9C4 FC
Brilliant Violet 650™ anti-human CD326 (EpCAM) 9C4 FC
Alexa Fluor® 594 anti-human CD326 (EpCAM) 9C4 IHC-P,ICC,SB
Purified anti-human CD326 (EpCAM) (Maxpar® Ready) 9C4 FC,CyTOF®
PE/Dazzle™ 594 anti-human CD326 (EpCAM) 9C4 FC
APC/Fire™ 750 anti-human CD326 (EpCAM) 9C4 FC
Brilliant Violet 510™ anti-human CD326 (EpCAM) 9C4 FC
Brilliant Violet 785™ anti-human CD326 (EpCAM) 9C4 FC
Brilliant Violet 711™ anti-human CD326 (Ep-CAM) 9C4 FC
TotalSeq™-A0123 anti-human CD326 (Ep-CAM) 9C4 PG
APC/Cyanine7 anti-human CD326 (Ep-CAM) 9C4 FC
Alexa Fluor® 700 anti-human CD326 (EpCAM) 9C4 FC
TotalSeq™-C0123 anti-human CD326 (Ep-CAM) 9C4 PG
TotalSeq™-B0123 anti-human CD326 (Ep-CAM) 9C4 PG
PE/Cyanine5 anti-human CD326 (Ep-CAM) 9C4 FC
TotalSeq™-D0123 anti-human CD326 (Ep-CAM) 9C4 PG
Spark UV™ 387 anti-human CD326 (Ep-CAM) 9C4 FC
Spark Red™ 718 anti-human CD326 (Ep-CAM) (Flexi-Fluor™) 9C4 FC
Spark Blue™ 574 anti-human CD326 (Ep-CAM) (Flexi-Fluor™) 9C4 FC
Spark Blue™ 550 anti-human CD326 (Ep-CAM) (Flexi-Fluor™) 9C4 FC
Go To Top Version: 3    Revision Date: 10.10.2024

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

 

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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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