Preparation and Storage
推奨アッセイ手順
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).
Product Notices
- 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.
- 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 Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
関連製品
The 6C3 monoclonal antibody specifically recognizes CD249 which is also known as, 6C3/BP-1, Enpep, or Ly-51. CD249 is a glycoprotein cell-surface differentiation antigen, which was originally identified on pre-B cell lymphomas (spontaneous and chemical- or retrovirus-transformed, in vitro and in vivo). CD249 is a homodimeric cell-surface glycoprotein with 140-kDa subunits that has been identified to possess aminopeptidase A (APA) activity. The same antigen is expressed at high levels on bone marrow stromal cell lines which support in vitro B lymphopoiesis, on thymic dendritic cells and cortical epithelial cells, and on a wide variety of mouse and rat tissues known to possess APA activity. Subsets of normal bone marrow pre-B and B lymphocytes express low levels of CD249 which is rapidly up-regulated on the pre-B cells in the presence of IL-7. A role for the CD249 molecule in the IL-7-driven proliferation of B cell precursors has been postulated. However, B-cell abnormalities were not detected in CD249/Ly-51-deficient mice. Mature B lymphocytes, thymocytes, peripheral T lymphocytes, erythroid cells, and myeloid cells (with the exception of thymic dendritic cells) do not express CD249. The 6C3 antibody can be used to identify cortical epithelium in frozen sections of thymuses from normal, SCID, and TCR-transgenic mice. It is possible that the low level of CD249 antigen detected, by flow cytometry, on some thymocytes may be passively adsorbed from adjacent epithelial cells during preparation of the cell suspensions.
The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max near 407 nm and Em Max near 421 nm, BD Horizon BV421 can be excited by the violet laser (405 nm) and detected with a 450/50 nm filter. BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue™ conjugates. Due to nearly identical excitation and emission properties but different spillover characteristics, BD Horizon BV421, Pacific Blue™, and BD Horizon V450 cannot be used simultaneously.
Development References (19)
-
Adkins B, Tidmarsh GF, Weissman IL. Normal thymic cortical epithelial cells developmentally regulate the expression of a B-lineage transformation-associated antigen. Immunity. 1988; 27(3):180-186. (Biology). View Reference
-
Goverman J, Brabb T, Huseby ES, Farr AG. TCR signaling regulates thymic organization: lessons from TCR-transgenic mice. Immunol Today. 1997; 18(5):204-208. (Biology). View Reference
-
Hardy RR, Carmack CE, Shinton SA, Kemp JD, Hayakawa K. Resolution and characterization of pro-B and pre-pro-B cell stages in normal mouse bone marrow. J Exp Med. 1991; 173(5):1213-1225. (Biology). View Reference
-
Li L, Wu Q, Wang J, Bucy RP, Cooper MD. Widespread tissue distribution of aminopeptidase A, an evolutionarily conserved ectoenzyme recognized by the BP-1 antibody. Tissue Antigens. 1993; 42(5):488-496. (Biology). View Reference
-
Lin Q, Taniuchi I, Kitamura D. T and B cell development in BP-1/6C3/aminopeptidase A-deficient mice. Int Immunol. 1998; 160(10):4681-4687. (Biology). View Reference
-
Morse HC 3rd, Tidmarsh GF, Holmes KL. Expression of the 6C3 antigen on murine hematopoietic neoplasms. Association with expression of abl, ras, fes, src, erbB, and Cas NS-1 oncogenes but not with myc. J Exp Med. 1987; 165(3):920-925. (Clone-specific). View Reference
-
Pillemer E, Whitlock C, Weissman IL. Transformation-associated proteins in murine B-cell lymphomas that are distinct from Abelson virus gene products. Proc Natl Acad Sci U S A. 1984; 81(14):4434-4438. (Immunogen). View Reference
-
Ramakrishnan L, Wu Q, Yue A, Cooper MD, Rosenberg N. BP-1/6C3 expression defines a differentiation stage of transformed pre-B cells and is not related to malignant potential. J Immunol. 1990; 145(5):1603-1608. (Biology). View Reference
-
Sherwood PJ, Weissman IL. The growth factor IL-7 induces expression of a transformation-associated antigen in normal pre-B cells. Int Immunol. 1990; 2(5):399-406. (Biology). View Reference
-
Surh CD, Ernst B, Sprent J. Growth of epithelial cells in the thymic medulla is under the control of mature T cells. J Exp Med. 1992; 172(2):611-616. (Biology). View Reference
-
Tidmarsh GF, Dailey MO, Whitlock CA, Pillemer E, Weissman IL. Transformed lymphocytes from Abelson-diseased mice express levels of a B lineage transformation-associated antigen elevated from that found on normal lymphocytes. J Exp Med. 1985; 162(5):1421-1434. (Clone-specific). View Reference
-
Vremec D, Shortman K. Dendritic cell subtypes in mouse lymphoid organs: cross-correlation of surface markers, changes with incubation, and differences among thymus, spleen, and lymph nodes. J Immunol. 1997; 159(2):565-573. (Biology). View Reference
-
Wang J, Cooper MD. Histidine residue in the zinc-binding motif of aminopeptidase A is critical for enzymatic activity. Proc Natl Acad Sci U S A. 1990; 90(4):1222-1226. (Biology). View Reference
-
Welch PA, Burrows PD, Namen A, Gillis S, Cooper MD. Bone marrow stromal cells and interleukin-7 induce coordinate expression of the BP-1/6C3 antigen and pre-B cell growth. Int Immunol. 1990; 2(8):697-705. (Biology). View Reference
-
Welch PA. Regulation of B cell precursor proliferation by aminopeptidase A. Int Immunol. 1995; 7(5):737-746. (Biology). View Reference
-
Wu L, Vremec D, Ardavin C, et al. Mouse thymus dendritic cells: kinetics of development and changes in surface markers during maturation. Eur J Immunol. 1995; 25(2):418-425. (Biology). View Reference
-
Wu Q, Lahti JM, Air GM, Burrows PD, Cooper MD. Molecular cloning of the murine BP-1/6C3 antigen: a member of the zinc-dependent metallopeptidase family. Proc Natl Acad Sci U S A. 1990; 87(3):993-997. (Biology). View Reference
-
Wu Q, Li L, Cooper MD, Pierres M, Gorvel JP. Aminopeptidase A activity of the murine B-lymphocyte differentiation antigen BP-1/6C3. Proc Natl Acad Sci U S A. 1991; 88(2):676-680. (Biology). View Reference
-
Wu Q, Tidmarsh GF, Welch PA, Pierce JH, Weissman IL, Cooper MD. The early B lineage antigen BP-1 and the transformation-associated antigen 6C3 are on the same molecule. J Immunol. 1989; 143(10):3303-3308. (Clone-specific). View Reference
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.
