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BB515 Mouse Anti-Human CD326
BB515 Mouse Anti-Human CD326
Flow cytometric analysis of CD326 expression on human SK-BR-3 cells. Cells from the human SK-BR-3 (Breast adenocarcinoma, ATCC HTB-30) cell line were stained with either BD Horizon™ BB515 Mouse IgG1, κ Isotype Control (Cat. No. 564416; dashed line histogram) or BD Horizon BB515 Mouse Anti-Human CD326 antibody (Cat. No. 565398; solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Flow cytometric analysis of CD326 expression on human SK-BR-3 cells. Cells from the human SK-BR-3 (Breast adenocarcinoma, ATCC HTB-30) cell line were stained with either BD Horizon™ BB515 Mouse IgG1, κ Isotype Control (Cat. No. 564416; dashed line histogram) or BD Horizon BB515 Mouse Anti-Human CD326 antibody (Cat. No. 565398; solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
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BD Horizon™
EPCAM; EGP; ESA; GA733-2; hEGP-2; KSA; M4S1; MIC18; MK-1; TACSTD1; TROP1
Human (QC Testing)
Mouse BALB/c IgG1, λ
Breast carcinoma–associated mucin BCA-225
Flow cytometry (Routinely Tested)
5 µl
4072
AB_2728107
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 with BD Horizon™ BB515 under optimum conditions and unconjugated antibody was removed.

Recommended Assay Procedures

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to CompBeads, they have spectral properties very similar to cells.   However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls.  It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

For optimal results, it is recommended to perform 2 washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescence staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.

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. The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. Diagnostic uses require a separate license.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
565398 Rev. 2
Antibody Details
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EBA-1

The EBA-1 monoclonal antibody specifically binds to human CD326. CD326 is an approximately 40 kDa type 1 transmembrane glycoprotein and adhesion molecule that mediates intercellular adhesive interactions. CD326 is also known as epithelial adhesion molecule (EpCAM), epithelial glycoprotein 2 (EGP-2), and epithelial surface antigen (ESA). The epithelial cells present in non-squamous epithelia and tumors derived from such cells show EpCAM expression. The normal epithelial cells reactive with anti-EpCAM antibodies are those present in the (lower) respiratory tract; the (lower) gastrointestinal tract; tubules in the kidney; the surface epithelium of the ovary; the exocrine and endocrine pancreas; secondary germ cells of telogenic hair follicles; and secretory tubules of sweat glands in the skin, whereas the epidermis is negative. In addition, all epithelial cells in the thyroid and epithelial cells in the thymus show EpCAM expression, while the outer cortex and Hassall's corpuscles have low expression. In the liver, only the bile ducts appear to be positive with anti-EpCAM antibodies. Non-squamous- carcinoma cells have high EpCAM expression; some squamous carcinoma cells. Tumors arising from non-epithelial cells, such as lymphoma, mesothelioma, neuroblastoma, and melanoma, do not express EpCAM.

The antibody was conjugated to BD Horizon BB515 which is part of the BD Horizon Brilliant™ Blue family of dyes. With an Ex Max near 490 nm and an Em Max near 515 nm, BD Horizon BB515 can be excited by the blue laser (488 nm) laser and detected with a 530/30 nm filter. This dye has been exclusively developed by BD Biosciences and is up to seven times brighter than FITC with less spillover into the PE channel. Due to similar excitation and emission properties, BB515, FITC, and Alexa Fluor® 488 cannot be used simultaneously. It is not recommended to use BB515 in cocktails that include Streptavidin conjugates as it may cause high background.

565398 Rev. 2
Format Details
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BB515
The BD Horizon Brilliant™ Blue 515 (BB515) dye is part of the BD Horizon Brilliant™ Blue family of dyes. This dye is a polymer fluorochrome with an excitation maximum (Ex Max) at 490-nm and an emission maximum (Em Max) of 515-nm. Driven by BD innovation, BB515 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 520-nm (e.g., 530/30-nm). BB515 reagents are significantly brighter than equivalent FITC or Alexa Fluor™ 488 reagents with less spillover into the PE detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
BB515
Blue 488 nm
490 nm
515 nm
565398 Rev.2
Citations & References
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Development References (13)

  1. Braun S, Pantel K, Müller P, et al. Cytokeratin-positive cells in the bone marrow and survival of patients with stage I, II, or III breast cancer. N Engl J Med. 2000; 342:525-533. (Biology). View Reference
  2. Carlsten M, Bjorkstrom NK, Norell H, et al. DNAX accessory molecule-1 mediated recognition of freshly isolated ovarian carcinoma by resting natural killer cells. Cancer Res. 2007; 67(3):1317-1325. (Clone-specific: Flow cytometry). View Reference
  3. De Leij L, Helrich W, Stein R, Mattes MJ. SCLC-cluster-2 antibodies detect the pancarcinoma/epithelial glycoprotein EGP-2. Int J Cancer. 1994; 8:60-63. (Biology). View Reference
  4. Diel IJ, Kaufmann M, Goerner R, Costa SD, Kaul S, Bastert G. Detection of tumor cells in bone marrow of patients with primary breast cancer: a prognostic factor for distant metastasis. J Clin Oncol. 1992; 10:1534-1539. (Biology). View Reference
  5. Hardingham JE, Kotasek D, Farmer B, et al. Immunobead-PCR: a technique for the detection of circulating tumor cells using immunomagnetic beads and the polymerase chain reaction. Cancer Res. 1993; 53(15):3455-3458. (Biology). View Reference
  6. Latza U, Niedobitek G, Schwarting R, Nekarda H, Stein H. Ber-EP4: new monoclonal antibody which distinguishes epithelia from mesothelial. J Clin Pathol. 1990; 43(3):213-219. (Biology). View Reference
  7. Momburg F, Moldenhauer G, Hämmerling GJ, Möller P. Immunohistochemical study of the expression of a Mr 34,000 human epithelium-specific surface glycoprotein in normal and malignant tissues. Cancer Res. 1987; 47:2883-2891. (Biology). View Reference
  8. Naume B, Borgen E, Beiske K, et al.. Immunomagnetic techniques for the enrichment and detection of isolated breast carcinoma cells in bone marrow and peripheral blood. J Hematother Stem Cell Res. 1997; 6:103-113. (Biology). View Reference
  9. Patriarca C, Macchi RM, Marschner AK, Mellstedt H. Epithelial cell adhesion molecule expression (CD326) in cancer: a short review. Cancer Treat Rev. 2012; 38(1):68-75. (Biology). View Reference
  10. Stahel RA, Gilks WR, Lehmann HP, Schenker T. Third International Workshop on Lung Tumor and Differentiation Antigens: overview of the results of the central data analysis. Int J Cancer. 1994; 8:6-26. (Biology). View Reference
  11. Takao M, Takeda K. Enumeration, characterization, and collection of intact circulating tumor cells by cross contamination-free flow cytometry. Cytometry A. 2011; 79(2):107-117. (Clone-specific: Flow cytometry). View Reference
  12. Trzpis M, McLaughlin PM, de Leij LM, Harmsen MC. Epithelial cell adhesion molecule: more than a carcinoma marker and adhesion molecule. Am J Pathol. 2007; 171(2):386-395. (Biology). View Reference
  13. Yemul S, Leon Ja, Pozniakoff T, Esser PD, Estabrook A. Radioimmunoimaging of human breast carcinoma xenografts in nude mouse model with 111In-labeled new monoclonal antibody EBA-1 and F(ab')2 fragments. Nucl Med Biol. 1993; 20:325-335. (Immunogen: ELISA, Radioimmunoassay, Western blot). View Reference
View All (13) View Less
565398 Rev. 2

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