Epcam g8.8

Manufactured by BD

Sourced in Germany

EpCAM (G8.8) is a laboratory reagent produced by BD. It is a monoclonal antibody that specifically binds to the epithelial cell adhesion molecule (EpCAM) protein.

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Lab products found in correlation

Foxp3 staining kit, by BD (1 mentions) Fixation permeabilization solution kit, by BD (1 mentions) Ter119 ter 119, by BioLegend (1 mentions) Taqman gene expression assay, by Thermo Fisher Scientific (1 mentions) Ki67 sola15, by Thermo Fisher Scientific (1 mentions) Efluor 780, by Thermo Fisher Scientific (1 mentions) Osteocalcin, by Abcam (1 mentions) Shandon cryomatrix, by Thermo Fisher Scientific (1 mentions) B220 ra3 6b2, by Thermo Fisher Scientific (1 mentions) Mac 1 m1 70, by Thermo Fisher Scientific (1 mentions) Trap kit, by Merck Group (1 mentions) Epr3776, by Abcam (1 mentions)

Close spelling variants of this product

EpCAM

3 protocols using epcam g8.8

Multiparametric Analysis of Immune Cells

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Mouse thymus, spleen, and lymph node (LN) were processed into single cell suspensions, and analyzed by flow cytometry. Thymic epithelial cells (TECs) were isolated as previously described (34 (link)). Dead cells were stained by a fixable viability dye conjugated to eFluor780 (eBioscience). Fixation and intracellular/intranuclear staining were performed using the eBioscience Foxp3 staining kit or BD Fixation/Permeabilization Solution Kit. The following antibodies were purchased from BD biosciences: TCRβ (H57–597), TCRγδ (GL-3) and EpCAM (G8.8). The following antibodies were purchased from eBioscience: CD3 (145–2C11), CD4 (GK1.5), CD11c (N418), CD25 (PC61.5), CD45R (B220, RA3–6B2), MHCII (M5/114) and Ki67 (SolA15). The following antibodies were purchased from BioLegend: CD8 (53–6.7), CD11b (M1/70), CD44 (IM7), CD45 (30-F11), CD62L (MEL-14), CD122 (TM-β1), Gr-1 (RB6–8C5), Ly-51 (6C3), NK-1.1 (PK136) and TER-119 (TER-119). UEA-1 was purchased from Vector Laboratories. Data were collected using a BD LSR II flow cytometer and were analyzed using FlowJo V10 (Treestar Inc.). The TaqMan Gene Expression Assay (ID: Mm00502000_m1, Thermo Fisher Scientific) for quantification of Klotho gene expression.

Xing Y., Smith M.J., Goetz C.A., McElmurry R.T., Parker S.L., Min D., Hollander G.A., Weinberg K.I., Tolar J., Stefanski H.E, & Blazar B.R. (2018). Thymic epithelial cell support of thymopoiesis does not require Klotho. Journal of immunology (Baltimore, Md. : 1950), 201(11), 3320-3328.

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Bone and Immune Cell Analysis

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Long bones were either embedded in Shandon cryomatrix (Thermo Fisher Scientific) or decalcified in 10% EDTA/PBS and embedded in paraffin. E14.5 FLs were embedded in cryomatrix. Cryosections were stained with tartrate-resistant acid phosphatase (TRAP kit; Sigma-Aldrich) and with antibodies against CD31 (R&D Systems), EpCAM (G8.8; BD), Osteocalcin (Abcam), and CD3e (145-2C11), B220 (RA3-6B2), Mac-1 (M1/70), and Gr-1 (RB6-8C5; eBioscience).

Ruppert R., Moser M., Sperandio M., Rognoni E., Orban M., Liu W.H., Schulz A.S., Oostendorp R.A., Massberg S, & Fässler R. (2015). Kindlin-3–mediated integrin adhesion is dispensable for quiescent but essential for activated hematopoietic stem cells. The Journal of Experimental Medicine, 212(9), 1415-1432.

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Immunohistochemical Assessment of EMT

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EpEX-specific (rat anti-mouse EpCAM G8.8, BD Biosciences, Heidelberg, Germany) and vimentin-specific antibodies (rabbit monoclonal to vimentin EPR3776, Abcam, Cambridge, UK) were used for IHC. Immunostaining was performed using the avidin-biotin-peroxidase method (VECTASTAIN, Vector Laboratories, Burlingame, CA, USA) according to the manufacturers’ protocol. IHC intensity scores (IHC score) were calculated as the product of intensity (0 to 3+) and the percentage of expressing tumor cells (score: 0 = 0 to 5%, 1 = 5 to 25%, 2 = 25 to 50%, 3 = 50 to 75%, and 4 = 75 to 100%). IHC scores represent averages of values independently assessed by a minimum of two experimenters, who were blinded to sample identity.
EMT scores were calculated as the percentage of mesenchymal, spindle-like cells (0 to 100%) and the level of cell-cell contact (disseminated cells represent 1 = 0 to 25%, 2 = 25 to 50%, 3 = 50 to 75%, and 4 = 75 to 100%). Three experimenters scored independently each cell line at an average confluence of 50 to 80% for three independent passages.

Liu X., Li J., Cadilha B.L., Markota A., Voigt C., Huang Z., Lin P.P., Wang D.D., Dai J., Kranz G., Krandick A., Libl D., Zitzelsberger H., Zagorski I., Braselmann H., Pan M., Zhu S., Huang Y., Niedermeyer S., Reichel C.A., Uhl B., Briukhovetska D., Suárez J., Kobold S., Gires O, & Wang H. (2019). Epithelial-type systemic breast carcinoma cells with a restricted mesenchymal transition are a major source of metastasis. Science Advances, 5(6), eaav4275.

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