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Purified Rat Anti-Mouse CD326
Purified Rat Anti-Mouse CD326

Multicolor analysis of the expression of Ep- CAM on splenic T lymphocytes. BALB/c splenocytes were stained with FITC Hamster Anti-Mouse CD3e (Cat. No 553061/553062), PE Rat Anti-Mouse CD25 (Cat. No. 553866), and either Purified Rat IgG2a, κ Isotype Control (Cat. No. 553927; Left Panel) or Purified Rat Anti-Mouse CD326 (Cat. No. 552370; Right Panel) in the presence of Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™; Cat. No. 553141/553142). The G8.8 and isotype control mAb were detected by Biotin Mouse anti-Rat IgG2a (Cat. No. 553894), followed by Streptavidin-APC (Cat. No. 554067). Viable lymphocytes were gated for exclusion of propidium iodide, and the CD25+ subpopulation was selected for analysis of CD3e and Ep-CAM expression. A small population of CD3+CD25+Ep-CAM+ cells were detected (Right Panel), whereas the CD25- T cells do not express Ep-CAM (data not shown). Flow cytometry was performed on a BD FACSCalibur™ (BD  Biosciences, San Jose, CA)

Multicolor analysis of the expression of Ep- CAM on splenic T lymphocytes. BALB/c splenocytes were stained with FITC Hamster Anti-Mouse CD3e (Cat. No 553061/553062), PE Rat Anti-Mouse CD25 (Cat. No. 553866), and either Purified Rat IgG2a, κ Isotype Control (Cat. No. 553927; Left Panel) or Purified Rat Anti-Mouse CD326 (Cat. No. 552370; Right Panel) in the presence of Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™; Cat. No. 553141/553142). The G8.8 and isotype control mAb were detected by Biotin Mouse anti-Rat IgG2a (Cat. No. 553894), followed by Streptavidin-APC (Cat. No. 554067). Viable lymphocytes were gated for exclusion of propidium iodide, and the CD25+ subpopulation was selected for analysis of CD3e and Ep-CAM expression. A small population of CD3+CD25+Ep-CAM+ cells were detected (Right Panel), whereas the CD25- T cells do not express Ep-CAM (data not shown). Flow cytometry was performed on a BD FACSCalibur™ (BD  Biosciences, San Jose, CA)

Product Details
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BD Pharmingen™
Ep-CAM; EGP; EGP-2; Egp314; GA733-2; TROP1; Tacsd1; Tacstd1; Ly74; gp40
Mouse (QC Testing)
Rat SD, also known as Sprague-Dawley (outbred) IgG2a, κ
Glycoconjugates from BALB/c mouse-derived TE-71 medullary thymic epithelial cell line
Flow cytometry (Routinely Tested), Immunohistochemistry-zinc-fixed (Tested During Development), Electron microscopy, Immunohistochemistry-frozen, Immunoprecipitation (Reported)
0.5 mg/ml
AB_394370
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Recommended Assay Procedures

Since this antigen is expressed at low density on the surface of thymocytes, it may be desirable to reduce background staining through the use of Purified Rat anti-Mouse CD16/CD32 (Mouse BD Fc Block™; Cat. No. 553142) and to amplify staining by using a biotinylated second-step antibody followed by a "bright" thirdstep reagent, such as Streptavidin-APC (Cat. No. 554067). If Mouse Fc Block is used, it is important the second-step antibody does not cross-react with the 2.4G2 mAb (rat IgG2b, κ).  We recommend Biotin Mouse anti-Rat IgG2a (Cat. No. 553894). Other reported applications include immunohistochemical staining of acetone-fixed frozen sections, zinc-fixed paraffin-embedded sections, immunoelectron microscopy, and immunoprecipitation.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  4. An isotype control should be used at the same concentration as the antibody of interest.
552370 Rev. 2
Antibody Details
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G8.8

The G8.8 monoclonal antibody recognizes CD326/Ep-CAM (Epithelial Cell Adhesion Molecule), also known as gp40 in the mouse and by a variety of names (including GA733-2, CO17-1A, and EGP) in the human.  In the mouse, Ep-CAM is a 40-42 kDa cell-surface type 1 transmembrane glycoprotein expressed on thymic epithelial cells, thymic dendritic cells, immature thymocytes, a small subset of peripheral T lymphocytes, intestinal epithelium, kidney-collecting tubule epithelium, keratinocytes, Langerhans cells and lymph node and splenic dendritic cells. Profiles of Ep-CAM expression on fetal thymocytes and on the CD4[-] CD8[-], CD4[+] CD8[+], CD4[-] CD8[+], and CD4[+] CD8[-] subsets of adult thymocytes have been published.  In unrelated studies, mouse Ep-CAM mRNA was detected in tissues containing epithelial cells (kidney, stomach, intestine, lung, and thymus) and in plasma cells and plasmacytomas, but not in heart, muscle, liver, brain, spleen, B lymphomas, or pre-B lymphomas.  Ep-CAM is a Ca[2+] independent homophilic adhesion molecule that is proposed to play roles in the development and normal function of epithelial tissues and in the progression of carcinomas.

552370 Rev. 2
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
552370 Rev.2
Citations & References
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Development References (9)

  1. Basak S, Speicher D, Eck S, Wunner W. Colorectal carcinoma invasion inhibition by CO17-1A/GA733 antigen and its murine homologue. J Natl Cancer Inst. 1998; 90(9):691-697. (Biology). View Reference
  2. Bergsagel PL, Victor-Kobrin C, Timblin CR, Trepel J, Kuehl WM. A murine cDNA encodes a pan-epithelial glycoprotein that is also expressed on plasma cells. J Immunol. 1992; 148(2):590-596. (Biology). View Reference
  3. Birebent B, Somasundaram R, Purev E. Anti-idiotypic antibody and recombinant antigen vaccines in colorectal cancer patients. Crit Rev Oncol Hematol. 2001; 39((1-2)):107-113. (Biology). View Reference
  4. Borkowski TA, Nelson AJ, Farr AG, Udey MC. Expression of gp40, the murine homologue of human epithelial cell adhesion molecule (Ep-CAM), by murine dendritic cells. Eur J Immunol. 1996; 26(1):110-114. (Clone-specific: Immunohistochemistry). View Reference
  5. Farr A, Nelson A, Truex J, Hosier S. Epithelial heterogeneity in the murine thymus: a cell surface glycoprotein expressed by subcapsular and medullary epithelium. J Histochem Cytochem. 1991; 39(5):345-353. (Immunogen: Electron microscopy, Immunohistochemistry, Immunoprecipitation). View Reference
  6. Litvinov SV, Balzar M, Winter MJ. Epithelial cell adhesion molecule (Ep-CAM) modulates cell-cell interactions mediated by classic cadherins. J Biol Chem. 1997; 139(5):1337-1348. (Biology). View Reference
  7. Nelson AJ, Dunn RJ, Peach R, Aruffo A, Farr AG. The murine homolog of human Ep-CAM, a homotypic adhesion molecule, is expressed by thymocytes and thymic epithelial cells. Eur J Immunol. 1996; 26(2):401-408. (Clone-specific: Electron microscopy, Immunohistochemistry, Immunoprecipitation). View Reference
  8. Taguchi N, Hashimoto Y, Naiki M. Abnormal thymic expression of epithelial cell adhesion molecule (EP-CAM) in New Zealand Black (NZB) mice. J Autoimmun. 1999; 13(4):393-400. (Clone-specific: Electron microscopy). View Reference
  9. Zutter MM. Gastrointestinal carcinoma antigen GA733: target for immunodestruction and potential modifier of invasiveness and chemoresponsiveness. J Natl Cancer Inst. 1998; 90(9):642-644. (Biology). View Reference
View All (9) View Less
552370 Rev. 2

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