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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 BD® 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 BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
Companion Products
The HMα5-1 antibody specifically recognizes the α5 subunit of the integrin α5β1 fibronectin receptor (CD49e/CD29, VLA-5) on mouse thymocytes and a variety of mouse cell lines, including pre-B and non-lymphoid cells, but not on mouse splenic or lymph node cells. It also detects rat CD49e on the RBL2H3 basophilic leukemia cell line,peritoneal mast cells, and endothelium, but not on splenocytes. It has been reported that soluble mAb HMα5-1 partially inhibits in vitro binding of a mouse cell line and of rat mast cells to fibronectin and inhibits the enhanced degranulation of IgE-sensitized rat RBL cells induced on fibronectin-coated plates. It has also been observed that plate-bound HMα5-1 antibody enhances the degranulation of IgEsensitized rat RBL-2H3 cells and that subcutaneous injection of HMα5-1 mAb into rats, along with anti-CD49d and anti-CD61 antibodies, inhibits experimentally induced passive cutaneous anaphylaxis.
Development References (4)
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Narumiya S, Abe Y, Kita Y, et al. Pre-B cells adhere to fibronectin via interactions of integrin alpha 5/alpha V with RGDS as well as of integrin alpha 4 with two distinct V region sequences at its different binding sites. Int Immunol. 1994; 6(1):139-147. (Immunogen). View Reference
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Noto K, Kato K, Okumura K, Yagita H. Identification and functional characterization of mouse CD29 with a mAb. Int Immunol. 1995; 7(5):835-842. (Clone-specific: Activation, Inhibition). View Reference
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Tanaka T, Ohtsuka Y, Yagita H, Shiratori Y, Omata M, Okumura K. Involvement of alpha 1 and alpha 4 integrins in gut mucosal injury of graft-versus-host disease. Int Immunol. 1995; 7(8):1183-1189. (Biology). View Reference
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Yasuda M, Hasunuma Y, Adachi H, et al. Expression and function of fibronectin binding integrins on rat mast cells.. Int Immunol. 1995; 7(2):251-8. (Immunogen: Activation, Immunoprecipitation, Inhibition). 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.