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- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036).
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- APC-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher.
- Cy is a trademark of GE Healthcare.
- APC-Cy7 is a tandem fluorochrome composed of Allophycocyanin (APC), which is excited by laser lines between 595 and 647 nm and serves as an energy donor, coupled to the cyanine dye Cy7™, which acts as an energy acceptor and fluoresces at 780 nm. BD Biosciences Pharmingen has maximized the fluorochrome energy transfer in APC-Cy7, thus maximizing its fluorescence emission intensity, minimizing residual emission from APC, and minimizing required electronic compensation in multilaser-laser flow cytometry systems. Note: Although every effort is made to minimize the lot-to-lot variation in residual emission from APC, it is strongly recommended that every lot be tested for differences in the amount of compensation required and that individual compensation controls are run for each APC-Cy7 conjugate.
- 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.
配套商品
The A20 monoclonal antibody specifically binds to CD45 (Leukocyte Common Antigen) on all leukocytes of mouse strains expressing the CD45.1 alloantigen (eg, RIII, SJL/J, STS/A, DA). This alloantigen was originally named Ly-5.2, but was later changed to Ly-5.1 to conform with the convention that the .2 alloantigen designates the C57BL/6 strain. mAb A20 has been reported not to react with leukocytes of most other mouse strains which express the CD45.2 alloantigen. CD45 is a member of the Protein Tyrosine Phosphatase (PTP) family; its intracellular (COOH-terminal) region contains two PTP catalytic domains while the extracellular region is highly variable due to alternative splicing of exons 4, 5, and 6 (designated A, B, and C, respectively), and differing levels of glycosylation. CD45 isoforms in the mouse are cell type-, maturation-, and activation state-specific. The CD45 isoforms play complex roles in T-cell and B-cell antigen receptor signal transduction. The A20 antibody has been reported to inhibit some responses of B cells (from mice expressing the CD45.1 alloantigen) to antigens and LPS.
研发参考 (7)
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Johnson P, Greenbaum L, Bottomly K, Trowbridge IS. Identification of the alternatively spliced exons of murine CD45 (T200) required for reactivity with B220 and other T200-restricted antibodies. J Exp Med. 1989; 169(3):1179-1184. (Biology). 查看参考
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Morse HC 3rd, Shen FW, Hammerling U. Genetic nomenclature for loci controlling mouse lymphocyte antigens. Immunogenetics. 1987; 25(2):71-78. (Biology). 查看参考
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Shen FW, Tung JS, Boyse EA. Further definition of the Ly-5 system. Immunogenetics. 1986; 24(3):146-149. (Biology). 查看参考
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Shen FW. Monoclonal antibodies to mouse lymphocyte differentiation alloantigens. In: Hammerling GJ, Hammerling U, Kearney JF, ed. Monoclonal Antibodies and T-cell Hybridomas; Perspectives and Technical Advances. 1981:25-31.
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Suzuki K, Oida T, Hamada H, et al. Gut cryptopatches: direct evidence of extrathymic anatomical sites for intestinal T lymphopoiesis. Immunity. 2000; 13(5):691-702. (Biology). 查看参考
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Yakura H, Kawabata I, Shen FW, Katagiri M. Selective inhibition of lipopolysaccharide-induced polyclonal IgG response by monoclonal Ly-5 antibody. J Immunol. 1986; 136(8):2729-2733. (Biology: Inhibition). 查看参考
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Yakura H, Shen FW, Bourcet E, Boyse EA. On the function of Ly-5 in the regulation of antigen-driven B cell differentiation. Comparison and contrast with Lyb-2. J Exp Med. 1983; 157(4):1077-1088. (Biology: Inhibition). 查看参考
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