Description
The 1F2 mAb reacts with Crry/p65, a cell-surface protein that blocks complement activation and C3 deposition by both the classical and alternative pathways. Based on nucleotide sequence identity, Crry/p65 is the genetic homologue of human CR1 (Complement Receptor 1). However, it has been proposed that Crry/p65 is the functional homologue of human MCP (Membrane Cofactor Protein) and/or DAF (Decay-Accelerating Factor), due to the similar functions these proteins share and the fact that Crry/p65 does not display CR1's complement-receptor activity. Crry/p65 is expressed in most organs, such as thymus, kidney, uterus, skin, lung, pancreas, heart, stomach, large and small intestine, and spleen. Platelets, brain, and muscle express low, but detectable levels of Crry/p65. In addition, Crry/p65 is expressed in J774 cells, bone marrow-derived macrophages, and thioglycollate-elicited peritoneal macrophages, as determined by RT-PCR. Crry is crucial in the regulation of complement activation during early mouse embryonic development and essential in fetomaternal tolerance. The 1F2 mAb partially blocks Crry/p65-mediated complement protection in vitro.
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.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions that minimize unconjugated dye and antibody.
1. 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.
2. Researchers should determine the optimal concentration of this reagent for their individual applications.
3. 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.
4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
5. 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.
6. An isotype control should be used at the same concentration as the antibody of interest.
7. 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.
8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
10. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
11. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
