|Presentation||Lyophilized from PBS, pH 7.4 containing 0.5% BSA and 0.1% sodium azide. Reconstitute with 1 mL of sterile distilled water. Concentration after reconstitution is 200 μg/mL.|
|Safety Information according to GHS|
|Material Size||200 µg|
|Reference overview||Pub Med ID|
|De novo generation of satellite DNA-based artificial chromosomes by induced large-scale amplification. |
Methods in molecular biology (Clifton, N.J.) 738 2011
Mammalian artificial chromosomes (MACs) are engineered chromosomes with defined genetic content that can function as non-integrating vectors with large carrying capacity and stability. The large carrying capacity allows the engineering of MACs with multiple copies of the same transgene, gene complexes, and to include regulatory elements necessary for the regulated expression of transgene(s). In recent years, different approaches have been explored to generate MACs (Vos Curr Opin Genet Dev 8:351-359, 1998; Danielle et al. Trends Biotech 23:573-583, 2005; Duncan and Hadlaczky Curr Opin Biotech 18:420-424, 2007): (1) the de novo formation by centromere seeding, the bottom-up approach, (2) the truncation of natural chromosomes or the modification of naturally occurring minichromosomes, the top-down approach, and (3) the in vivo inductive approach. Satellite DNA-based artificial chromosomes (SATACs) generated by the in vivo inductive method have the potential to become an efficient tool in diverse gene technology applications such as cellular protein manufacturing (Kennard et al. BioPharm Int 20:52-59, 2007; Kennard et al. Biotechnol Bioeng 104:526-539, 2009; Kennard et al. Biotechnol Bioeng 104:540-553, 2009), transgenic animal production (Telenius et al. Chromosome Res 7:3-7, 1999; Co et al. Chromosome Res 8:183-191, 2000; Monteith et al. Methods Mol Biol 240:227-242, 2003), and ultimately a safe vector for gene therapy (Vanderbyl et al. Stem Cells 22:324-333, 2004; Vanderbyl et al. Exp Hematol 33:1470-1476, 2005; Katona et al. Cell. Mol. Life Sci 65:3830-3838, 2008). A detailed protocol for the de novo generation of satellite DNA-based artificial chromosomes (SATACs) via induced large-scale amplification is presented.
|Monoclonal antibodies to a nuclear protein (PCNA/cyclin) associated with DNA replication. |
Ogata, K, et al.
Exp. Cell Res., 168: 475-86 (1987) 1987
Two hybridomas producing monoclonal antibodies to proliferating cell nuclear antigen. (PNCA)/cyclin were generated from spleen cells of BALB/c mice immunized with purified PCNA from rabbit thymus. The specificity of the monoclonal antibodies (19A2 and 19F4) was established by showing that they reacted in enzyme-linked immunosorbent assay (ELISA) with purified PCNA. Furthermore, they reacted in one-dimensional (ID) gel immunoblots with a 36 kD polypeptide which also reacted with human autoantibodies from lupus patients. Both monoclonals also recognized the nuclear polypeptide cyclin in two-dimensional (2D) gel immunoblots of HeLa cell proteins. Epitopes recognized by 19A2 and 19F4 were analysed by competitive inhibition test using a modified ELISA. The data suggested that the epitopes were closely related, but not identical. The data with human auto-antibodies were more difficult to interpret, although it suggested that some human anti-PCNA may share epitopes with 19A2 and 19F4, but in addition recognize different epitopes on the PCNA molecule.
|Detection of plasma cell immunoglobulins in tissue sections optimally fixed for ultrastructural immunocytochemistry. |
Gonatas, N K, et al.
J. Histochem. Cytochem., 35: 189-96 (1987) 1987
We describe the ultrastructural localization of plasma cell immunoglobulins in vibratome sections of popliteal lymph nodes. Fixation with glutaraldehyde-paraformaldehyde gave better tissue and antigen preservation than paraformaldehyde or periodic acid lysine-paraformaldehyde; biotinylated Fab fragments of sheep anti-mouse IgG-streptavidin-biotinylated horseradish peroxidase (HRP) or Fab-HRP conjugates gave similar results. With both immunoreagents, excellent tissue preservation and antigen detection was observed in the first layer of cells sectioned with the vibratome. Conjugates of anti-mouse IgG with HRP did not show any staining. Peroxidase stain was observed in the nuclear envelope, cisternae of the rough endoplasmic reticulum, and the Golgi apparatus complex. In the Golgi apparatus, staining was seen consistently in cisternae of the cis face and in adjacent vesicles; the trans cisternae showed weak or no stain, and adjacent vesicles, "coated" vesicles, and granules were not stained. This study shows that high quality of tissue preservation and antigen detection, by both light and ultrastructural immunocytochemistry, is feasible in tissue fixed with glutaraldehyde-paraformaldehyde followed by vibratome sectioning and immunostaining with Fab-biotin-streptavidin-biotin-HRP, or Fab-HRP.
|A cell ELISA for the quantitation of leukocyte antigens. Requirements for calibration. |
J. Immunol. Methods, 94: 91-8 (1986) 1986
|Utilization of the biotin/avidin system to amplify the sensitivity of the enzyme-linked immunosorbent assay (ELISA). |
Kendall, C, et al.
J. Immunol. Methods, 56: 329-39 (1983) 1983
The biotin/avidin system was incorporated into the enzyme-linked immunosorbent assay (ELISA) technique to increase the sensitivity of the standard ELISA for the detection of mouse antibody to hepatitis B surface antigen ((anti-HBs and HBsAg, respectively). Two biotin/avidin ELISA designs were studied. In both assays, 96-well polystyrene plates were coated with HBsAg, post-coated with 0.5% gelatin and incubated with dilutions of mouse anti-HBs. In the biotin/avidin (BA) ELISA, reagents were added to antibody reacted wells in the following sequence: biotinylated goat anti-mouse IgG (b-GAMG), avidin-alkaline phosphatase (Av-AP) and substrate. The order of reactants after mouse antibody in the biotin/avidin/biotin (BAB) ELISA was b-GAMG, avidin, biotinylated alkaline phosphatase (b-AP) and substrate. The sensitivities of BA ELISA, BAB ELISA and a standard ELISA using a glutaraldehyde conjugated goat anti-mouse enzyme were compared to AUSAB (a commercial radioimmunoassay) using a panel of 23 mouse anti-HBs sera. All 3 ELISAs were more sensitive than AUSAB; the standard ELISA, BAB ELISA and BA ELISA were respectively 50, 1173 and 4134 times more sensitive than AUSAB for detection of mouse anti-HBs activity.
|Sheep anti-Mouse Ig, (H+L) Digoxigenin Conjugated Affinity Purified F(ab')2 antibody - Data Sheet|