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BV421 Rat Anti-Mouse CD326
BV421 Rat Anti-Mouse CD326
Multicolor 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 BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Cat. No. 562602) or BD Horizon™ BV421 Rat Anti-Mouse CD326 antibody (Cat. No. 563214).    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) and Ig Isotype Control (dashed line) 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 FITC Hamster Anti-Mouse CD3e (Cat. No 553062/553061/561827) and PE Rat Anti-Mouse CD25 (Cat. No. 553866/561065) antibodies. Two-color flow cytometric dot plots show the correlated expression patterns of Ig Isotype control staining (Middle Plot) or CD326 (Right Plot) 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.
Multicolor 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 BD Horizon™ BV421 Rat IgG2a, κ Isotype Control (Cat. No. 562602) or BD Horizon™ BV421 Rat Anti-Mouse CD326 antibody (Cat. No. 563214).    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) and Ig Isotype Control (dashed line) 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 FITC Hamster Anti-Mouse CD3e (Cat. No 553062/553061/561827) and PE Rat Anti-Mouse CD25 (Cat. No. 553866/561065) antibodies. Two-color flow cytometric dot plots show the correlated expression patterns of Ig Isotype control staining (Middle Plot) or CD326 (Right Plot) 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 Horizon™
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
17075
AB_2738073
Aqueous buffered solution containing BSA and ≤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 with BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  5. 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.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  8. Brilliant Violet™ 421 is a trademark of Sirigen.
563214 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.

The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon™ Brilliant Violet™ family of dyes.  With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon™ BV421  can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon™ BV421  conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue™ conjugates.

563214 Rev. 1
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
563214 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
563214 Rev. 1

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