Preparation And Storage
Recommended Assay Procedures
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 or 566349).
When setting up compensation, it is recommended to compare spillover values obtained from cells and BD™ CompBeads to ensure that beads will provide sufficiently accurate spillover values.
For optimal results, it is recommended to perform two washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescent 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
- This antibody was developed for use in flow cytometry.
- The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- BD Horizon Brilliant Blue 700 is covered by one or more of the following US patents: 8,455,613 and 8,575,303.
- Cy is a trademark of GE Healthcare.
Companion Products
KS1/4 antigen (Ag), defined by the KS1/4 antibody (KS1/4), is expressed in many epithelial-derived carcinomas and normal epithelial cell surfaces. The cDNA encoding KS1/4 has been isolated and contains an open reading frame of 314 amino acids including a putative signal sequence. KS1/4 Ag migrates as three glycosylated polypeptides with molecular masses of 35, 40 and 42 kDa. The 40 and 42 kDa species are similar proteins and appear to differ only by their degree of N-linked glycosylation. The 35 kDa species results from proteolytic cleavages of the larger molecular weight proteins. KS1/4 was first described as a monoclonal antibody which recognized a lung adenocarcinoma associated antigen. In oncolytic drug targeting studies, KS1/4 suppressed the growth of human lung tumor xenografts in athymic nude mice. Subsequent studies showed that KS1/4 reacts with a variety of tumor tissues including colon, breast, ovarian, and pancreas. It also recognizes normal epithelium including colon, stomach, small intestine, liver, kidney, lung, pancreas, skin and ovary. These results suggest that KS1/4 Ag represents an epithelial cell/epithelial-derived carcinoma marker. KS1/4 recognizes a 40 - 42 kDa antigen expressed on the cell surfaces of a variety of epithelial tumors and normal epithelial cell types. KS1/4 also recognizes a 35 kDa proteolytic fragment. UCLA-P3 cells derived from a human adenocarcinoma of the lung were used as immunogen.
The antibody was conjugated to BD Horizon™ BB700, which is part of the BD Horizon Brilliant™ Blue family of dyes. It is a polymer-based tandem dye developed exclusively by BD Biosciences. With an excitation max of 485 nm and an emission max of 693 nm, BD Horizon BB700 can be excited by the 488 nm laser and detected in a standard PerCP-Cy™5.5 set (eg, 695/40-nm filter). This dye provides a much brighter alternative to PerCP-Cy5.5 with less cross laser excitation off the 405 nm and 355 nm lasers.
Development References (5)
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Bumol TF, Marder P, DeHerdt SV, Borowitz MJ, Apelgren LD. Characterization of the human tumor and normal tissue reactivity of the KS1/4 monoclonal antibody. Hybridoma. 1988; 7(4):407-415. (Clone-specific: Flow cytometry, Immunohistochemistry). View Reference
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Perez MS, Walker LE. Isolation and characterization of a cDNA encoding the KS1/4 epithelial carcinoma marker. J Immunol. 1989; 142(10):3662-3667. (Clone-specific: Immunoprecipitation). View Reference
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Spearman ME, Goodwin RM, Apelgren LD, Bumol TF. Disposition of the monoclonal antibody-vinca alkaloid conjugate KS1/4-DAVLB (LY256787) and free 4-desacetylvinblastine in tumor-bearing nude mice. J Pharmacol Exp Ther. 1987; 241(2):695-703. (Biology). View Reference
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Varki NM, Reisfeld RA, Walker LE. Antigens associated with a human lung adenocarcinoma defined by monoclonal antibodies. Cancer Res. 1984; 44(2):681-687. (Immunogen: Immunofluorescence, Immunohistochemistry, Immunoprecipitation). View Reference
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Varki NM, Reisfeld RA, Walker LE. Effect of monoclonal antibody-drug conjugates in teh in vivody growth of h uman tumors established in nude mice. In: Reisfeld R, Sell S, ed. Monoclonal Antibodies in Cancer Therapy, vol. 27. New York: Alan R Liss, Inc; 195:207.
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
