Preparation And Storage
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
- 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).
- 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.
- 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.
Companion Products
The C40-1457 monoclonal antibody specifically recognizes CD271 that is also known as the nerve growth factor receptor (NGFR). CD271 is 75 kDa type I transmembrane glycoprotein likewise known as TNFRSF16 that belongs to the tumor necrosis factor receptor (TNFR) superfamily. CD271 has been found localized to neuronal axons, Schwann cells, and perineural cells of peripheral nerves. It is also expressed by some epithelial, mesenchymal and lymphoid tissues. NGFR is the receptor for nerve growth factor (NGF), a polypeptide that is essential for normal development of the nervous system. NGF promotes survival and differentiation of sympathetic and sensory neurons during embryological development of the peripheral nervous system. NGF binds to two distinctive surface receptors, the p/140[prototrk] and p75[NGFR]. High affinity binding of NGF requires that both receptor molecules be expressed. NGFR is expressed on human and rat lymphocytes. A subset of lymphoid cells in the spleen, lymph nodes, and follicular dendritic cells in germinal centers of reactive lymph nodes were found to express CD271. It has been reported that NGFR interaction with its ligand, NGF, may play a role in immunoregulation. NGF may also function as a B-cell growth factor.
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.
Development References (7)
-
Brodie C, Gelfand EW. Functional nerve growth factor receptors on human B lymphocytes. Interaction with IL-2. J Immunol. 1992; 148(11):3492-3497. (Biology). View Reference
-
Chesa PG, Rettig WJ, Thomson TM, Old LJ, Melamed MR. Immunohistochemical analysis of nerve growth factor receptor expression in normal and malignant human tissues. J Histochem Cytochem. 1988; 36(4):383-389. (Biology). View Reference
-
Hempstead BL, Martin-Zanca D, Kaplan DR, Parada LF, Chao MV. High-affinity NGF binding requires coexpression of the trk proto-oncogene and the low-affinity NGF receptor. Nature. 1991; 350(6320):678-683. (Biology). View Reference
-
Holling TM, Bergevoet MW, Wilson L, et al. A Role for EZH2 in Silencing of IFN-γ Inducible MHC2TA Transcription in Uveal Melanoma. J Immunol. 2007; 179(8):5317-5325. (Biology). View Reference
-
Kanellopoulou C, Muljo SA, Dimitrov SD, et al. X chromosome inactivation in the absence of Dicer.. Proc Natl Acad Sci U S A. 2009; 106(4):1122-1127. (Biology). View Reference
-
Thompson SJ, Schatteman GC, Gown AM, Bothwell M. A monoclonal antibody against nerve growth factor receptor. Immunohistochemical analysis of normal and neoplastic human tissue. Am J Clin Pathol. 1989; 92(4):415-423. (Biology). View Reference
-
Van Landuyt K, Lories R, Jones E, et al. Flow cytometric characterization of freshly isolated and culture expanded human synovial cell populations in patients with chronic arthritis. Arthritis Res Ther. 2001; 12(1):R15. (Clone-specific: Flow cytometry). 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.