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APC Rat Anti-Mouse CD326
APC Rat Anti-Mouse CD326

Two-color analysis of CD326 expression on mouse thymocytes and splenic T lymphocytes. BALB/c mouse thymocytes and splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142) and stained with APC Rat IgG2a, κ Isotype Control (Cat. No. 554690) or APC Rat Anti-Mouse CD326 antibody (Cat. No. 563478).

     Left Panel: Thymocytes were further stained with FITC Rat Anti-Mouse CD4 (553047/553046/561835) and PE Rat Anti-Mouse CD8a (553033/553032/561095) antibodies. The CD326 (solid line histogram) and Ig Isotype Control (dashed line histogram) fluorescence histograms were derived from CD4- and CD8-negative gated events with the forward and side light-scatter characteristics of viable thymocytes.

      Middle and Right Panels: The splenic leucocytes were further stained with PE Hamster Anti-Mouse CD3e (Cat. No. 553064/553063/561824) and BD Horizon™ PE-CF594 Rat Anti-Mouse CD25 (Cat. No. 562694/562695) antibodies. Two-color flow cytometric dot plots show the correlated expression patterns of Ig Isotype control staining (Middle Panel) or CD326 (Right Panel) versus CD3e for CD25+ gated events with the forward and side light-scatter characteristics of viable splenic leucocytes. A small population of CD3+CD25+CD326+ cells were detected (Right Panel), whereas the CD25- T cells do not express detectable levels of CD326 (data not shown).

      Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Two-color analysis of CD326 expression on mouse thymocytes and splenic T lymphocytes. BALB/c mouse thymocytes and splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142) and stained with APC Rat IgG2a, κ Isotype Control (Cat. No. 554690) or APC Rat Anti-Mouse CD326 antibody (Cat. No. 563478).

     Left Panel: Thymocytes were further stained with FITC Rat Anti-Mouse CD4 (553047/553046/561835) and PE Rat Anti-Mouse CD8a (553033/553032/561095) antibodies. The CD326 (solid line histogram) and Ig Isotype Control (dashed line histogram) fluorescence histograms were derived from CD4- and CD8-negative gated events with the forward and side light-scatter characteristics of viable thymocytes.

      Middle and Right Panels: The splenic leucocytes were further stained with PE Hamster Anti-Mouse CD3e (Cat. No. 553064/553063/561824) and BD Horizon™ PE-CF594 Rat Anti-Mouse CD25 (Cat. No. 562694/562695) antibodies. Two-color flow cytometric dot plots show the correlated expression patterns of Ig Isotype control staining (Middle Panel) or CD326 (Right Panel) versus CD3e for CD25+ gated events with the forward and side light-scatter characteristics of viable splenic leucocytes. A small population of CD3+CD25+CD326+ cells were detected (Right Panel), whereas the CD25- T cells do not express detectable levels of CD326 (data not shown).

      Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

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)
0.2 mg/ml
AB_2738234
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to APC under optimum conditions, and unconjugated antibody and free APC were removed.

Product Notices

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  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. This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
  5. CF™ is a trademark of Biotium, Inc.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
563478 Rev. 1
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.

563478 Rev. 1
Format Details
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APC
Allophycocyanin (APC), is part of the BD family of phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 651 nm and an emission maximum (Em Max) at 660 nm. APC is designed to be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 660 nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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APC
Red 627-640 nm
651 nm
660 nm
563478 Rev.1
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
563478 Rev. 1

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