Alexa Fluor® 647 anti-mouse CD326 (Ep-CAM) Antibody

Product Details

Verified Reactivity

Mouse

Antibody Type

Monoclonal

Host Species

Rat

Immunogen

TE-71 thymic epithelial cell line

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® 647 under optimal conditions.

Concentration

0.5 mg/ml

Storage & Handling

The CD326 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
IHC-F, 3D IHC - Verified

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 ≤ 0.25 µg per 10⁶ cells in 100 µl volume. For immunohistochemistry on frozen tissue sections, a concentration range of 2.5 - 5.0 µg/ml is suggested. For 3D immunohistochemistry on formalin-fixed tissues, a concentration of 5.0 µg/mL is suggested. It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 647 has a maximum emission of 668 nm when it is excited at 633nm / 635nm.

Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.

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Excitation Laser

Red Laser (633 nm)

Application Notes

Additional reported applications for clone G8.8 (for the relevant formats) include: immunohistochemistry of frozen sections: acetone fixed¹, with or without OCT embedding^2,4, and spatial biology (IBEX)^13,14.

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. Farr A, et al. 1991. J. Histochem. Cytochem. 39:645. (FC, IHC)
2. Dooley J, et al. 2005. J. Immunol. 175:4331. (FC, IHC)
3. Hinterberger M, et. al. 2010. Nat. Immunol. 11:512. (FC) PubMed
4. Gracz AD, et al. 2010. Am J. Physiol Gastrointest Liver Physiol. 298:590. (IHC) PubMed
5. Nudel I, et al. 2011. J. Immunol. 186:891. PubMed
6. Morimoto H, et al. 2012. Biol Reprod. 86:148. PubMed
7. Ishii K, et al. 2012. Development. 139:1734. PubMed
8. Takehashi M, et al. 2012. Biol Reprod. 86:178. PubMed
9. Murakami R, et al. 2013. PLoS One. 8:73270. PubMed
10. Taguchi K, et al. 2014. Mol Cell Biol. 34:900. PubMed
11. Hirokawa Y, et al. 2014. Am J Physiol Gastrointerest Liver Physiol. 306:547. PubMed
12. Ding X, et al. 2015. Cancer Res. 75:330. PubMed
13. Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed
14. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed

Product Citations

1. Gabitova-Cornell L, et al. 2020. Cancer Cell. 38(4):567-583.e11. PubMed
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4. Moskwa N, et al. 2022. Development. 149:. PubMed
5. Mueller JPJ, et al. 2023. Front Immunol. 14:1034032. PubMed
6. Bilate AM, et al. 2020. Immunity. 53(5):1001-1014.e20. PubMed
7. Davis F, et al. 2015. Proc Natl Acad Sci U S A. 112:5827. PubMed
8. Molaro A, et al. 2014. Genes Dev. 28:1544. PubMed
9. Biffi G, et al. 2018. Cancer Discov. 2:282. PubMed
10. McClendon J, et al. 2017. Am J Pathol. 10.1016/j.ajpath.2017.04.012. PubMed
11. Strunz M, et al. 2020. Nat Commun. 2.929861111. PubMed
12. Blijswijk J, et al. 2015. J Immunol. 194:307. PubMed
13. Lin JR et al. 2018. eLife. 7 pii: e31657. PubMed
14. Riemondy KA, et al. 2019. JCI Insight. 5:. PubMed
15. Joshi PA, et al. 2019. Nat Commun. 10:1760. PubMed
16. Burns KA, et al. 2018. Endocrinology. 159:103. PubMed
17. Paiva RA, et al. 2021. Cell Reports. 35(2):108967. PubMed
18. Cheng Q, et al. 2020. Nat Nanotechnol. 0.842361111. PubMed
19. Progatzky F, et al. 2021. Nature. 599:125. PubMed
20. Xiong H, et al. 2021. Theranostics. 11:1594. PubMed
21. Cohen SB et al. 2018. Cell host & microbe. 24(3):439-446 . PubMed
22. Campbell C et al. 2018. Immunity. 48(6):1245-1257 . PubMed
23. Zhang J, et al. 2018. Sci Rep. 8:2373. PubMed
24. Dravis C et al. 2018. Cancer cell. 34(3):466-482 . PubMed
25. Moon S, et al. 2021. J Exp Med. 218:. PubMed
26. Madenspacher JH, et al. 2020. JCI Insight. 5:00. PubMed
27. Heijden M, et al. 2016. Nat Commun. 7:10916. PubMed
28. Shaffer M, et al. 2010. PLoS One. 5:e12404. PubMed
29. Hegab A, et al. 2015. Stem Cell Res. 15: 109-121. PubMed
30. Kolodkin-Gal D, et al. 2021. Gut. Online ahead of print. PubMed
31. Ma Z, et al. 2022. Methods Mol Biol. 2471:49. PubMed
32. Tang R, et al. 2020. Elife. 9:00. PubMed
33. Giraddi R, et al. 2015. Nat Commun. 6: 8487. PubMed
34. Bernstock JD, et al. 2019. J Allergy Clin Immunol. 145:358. PubMed
35. Stewart AS, et al. 2021. Am J Physiol Gastrointest Liver Physiol. 321:G588. PubMed
36. Steele NG, et al. 2021. Clin Cancer Res. 27:2023. PubMed
37. Wasik K, et al. 2015. Genes Dev. 29: 1403 - 1415. PubMed
38. Miragaia RJ, et al. 2018. Sci Rep. 8:685. PubMed
39. Satoh–Takayama N, et al. 2020. Immunity. 52(4):635-649. PubMed
40. Fang Y, et al. 2022. Development. 149:. PubMed
41. Kurashima Y, et al. 2021. Nat Commun. 12:1067. PubMed
42. Liu Y, et al. 2020. Cell. 183(6):1665-1681.e18. PubMed
43. Gaiser M, et al. 2012. Proc Natl Acad Sci U S A. 1.034027778. PubMed
44. Giraddi RR et al. 2018. Cell reports. 24(6):1653-1666 . PubMed
45. Koyama M et al. 2019. Immunity. 51(5):885-898 . PubMed
46. Bretz N, et al. 2014. Immunol Lett. 161:140. PubMed
47. Oni TE, et al. 2020. J Exp Med. :217. PubMed
48. Chung CY et al. 2019. Cell Rep. 29(2):495-510 . PubMed

RRID

AB_1134104 (BioLegend Cat. No. 118211)
AB_1134104 (BioLegend Cat. No. 118212)

Antigen Details

Structure

40 kD single-pass type 1 glycoprotein. 293 amino acids, with a 21 aa signal peptide, a 246 aa extracellular domain, a 21 aa transmembrane domain, and a 26 aa cytoplasmic domain. The extracellular domain contains two epidermal growth factor-like repeats.

Distribution

Expressed on majority of epithelial cell membranes with the exception of adult squamous cells of the skin and a few specific epithelial cell types.

Function

Mediates calcium-independent homophilic cell-cell adhesion.

Interaction

CD326 displays hemophilic binding.

Ligand/Receptor

CD305 (LAIR-1), CD306 (LAIR-2), and Ep-CAM.

Cell Type

Embryonic Stem Cells, Epithelial cells

Biology Area

Immunology, Stem Cells

Molecular Family

Adhesion Molecules, CD Molecules

Antigen References

1. Borkowski TA, et al. 1996. Eur. J. Immunol. 26:110.
2. Bergsagel PL, et al. 1992. J. Immunol. 148:590.

Gene ID

17075 View all products for this Gene ID

UniProt

View information about CD326 on UniProt.org

Related Protocols

• Cell Surface Flow Cytometry Staining Protocol
• Ce3D™ Tissue Clearing Kit

Other Formats