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Alexa Fluor® 488 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
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Product Citations
publications
1_9C4_Alx488_020708
Human colon carcinoma cell line HT29 stained with 9C4 Alexa Fluor® 488
  • 1_9C4_Alx488_020708
    Human colon carcinoma cell line HT29 stained with 9C4 Alexa Fluor® 488
  • 2_24_Human_Jejunum_EpCAM_CD49a
    Confocal image of human jejunum sample acquired using the IBEX method of highly multiplexed antibody-based imaging: EpCAM (purple) in Cycle 1 and CD49a (cyan) in Cycle 2. Tissues were prepared using ~1% (vol/vol) formaldehyde and a detergent. Following fixation, samples are immersed in 30% (wt/vol) sucrose for cryoprotection. Images are courtesy of Drs. Andrea J. Radtke and Ronald N. Germain of the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
  • 3_28_Human_Jejunum_EpCAM_CD44
    Confocal image of human jejunum sample acquired using the IBEX method of highly multiplexed antibody-based imaging: EpCAM (blue) in Cycle 1 and CD44 (yellow) in Cycle 6. Tissues were prepared using ~1% (vol/vol) formaldehyde and a detergent. Following fixation, samples are immersed in 30% (wt/vol) sucrose for cryoprotection. Images are courtesy of Drs. Andrea J. Radtke and Ronald N. Germain of the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
See Alexa Fluor® 488 spectral data
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324209 25 tests 113€
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324210 100 tests 235€
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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 (pdf)

Product Details

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 Alexa Fluor® 488 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 - Reported in the literature, not verified in house
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. 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

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

Additional Product Notes

Iterative Bleaching Extended multi-pleXity (IBEX) is a fluorescent imaging technique capable of highly-multiplexed spatial analysis. The method relies on cyclical bleaching of panels of fluorescent antibodies in order to image and analyze many markers over multiple cycles of staining, imaging, and, bleaching. It is a community-developed open-access method developed by the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).

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. Cai X, et al. 2017. Neoplasia. 10.1016/j.neo.2017.03.005. PubMed
  2. Gell JJ, et al. 2018. Stem Cell Res. 27:46. PubMed
  3. Hu H et al. 2018. Cell. 175(6):1591-1606 . PubMed
  4. Chen D et al. 2018. Cell reports. 25(13):3591-3602 . PubMed
  5. Chen D, et al. 2017. Biol Reprod. 97:850. PubMed
  6. Forbes TA, et al. 2018. Am J Hum Genet. 102:816. PubMed
  7. Koyama M et al. 2019. Immunity. 51(5):885-898 . PubMed
  8. Pal D et al. 2017. eLife. 6 pii: e21615. PubMed
  9. Gerdur ísberg ó, et al. 2019. Sci Rep. 9:14843. PubMed
  10. Chen D, et al. 2019. Cell Rep. 29:4568. PubMed
  11. Calandrini C, et al. 2020. Nat Commun. 11:1310. PubMed
  12. Vanslambrouck JM, et al. 2019. J Am Soc Nephrol. 30:1811. PubMed
  13. Hong HK, et al. 2019. Oncol Rep. 42:2029. PubMed
  14. Horwitz K, et al. 2008. Proc Natl Acad Sci U S A. 105:5774. PubMed
  15. Morris K, et al. 2014. PLoS One. 9:98656. PubMed
  16. Noort V, et al. 2014. Cancer Res. 74:5690. PubMed
  17. Cai X, et al. 2014. PLoS One. 9:108942. PubMed
  18. Catenacci D, et al. 2015. Gastroenterology. 149: 1794-1803.e4. PubMed
  19. Morsing M, et al. 2020. Breast Cancer Res. 0.9875. PubMed
  20. Zabala M, et al. 2020. Cell Stem Cell. 27(2):284-299.e8. PubMed
  21. Yu L, et al. 2020. Cell Stem Cell. 28(3):550-567.e12. PubMed
  22. Howden SE, et al. 2020. Cell Stem Cell. 28(4):671-684.e6. PubMed
  23. Mourcin F, et al. 2021. Immunity. . PubMed
  24. Cardenas JJ, et al. 2020. Methods Enzymol. 415:631. PubMed
  25. Pyo DH, et al. 2020. Cancer Biol Ther. 495:21. PubMed
  26. Chitiashvili T, et al. 2020. Nat Cell Biol. 22:1436. PubMed
  27. Safaric Tepes P, et al. 2021. Elife. 10: . PubMed
  28. Sullivan NT, et al. 2022. STAR Protoc. 3:101367. PubMed
  29. Staudte S, et al. 2022. Cancers (Basel). 14:. PubMed
  30. Carraro G, et al. 2021. Nat Med. 27:806. PubMed
  31. Palkowitz AL, et al. 2021. Adv Healthc Mater. 10:e2100132. PubMed
  32. Hancock GV, et al. 2021. Stem Cell Res. 55:102493. PubMed
  33. Inde Z, et al. 2021. Sci Adv. 7:. PubMed
  34. Sandlin CW, et al. 2022. PLoS One. 17:e0274091. PubMed
  35. Vanslambrouck JM, et al. 2022. Nat Commun. 13:5943. PubMed
  36. Pandolfi EC, et al. 2022. Cell Rep Med. 3:100782. PubMed
  37. Jeong HO, et al. 2022. iScience. 25:105358. PubMed
  38. Qin XY, et al. 2023. Cell Death Dis. 14:358. PubMed
RRID
AB_756083 (BioLegend Cat. No. 324209)
AB_756084 (BioLegend Cat. No. 324210)

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

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: 4    Revision Date: 04.18.2022

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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.
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