To detect human glioblastoma cells in mouse brain, we used a 1:100 dilution of the mouse anti-vimentin antibody clone V9 (Dako, M0725), which has no cross-reactivity with mouse vimentin (Valadez et al., 2014 ). We also determined the levels of immune cells positive for CD3-Ɛ using a 1:10 dilution of Armenian Hamster anti-CD3-Ɛ antibody-FITC followed by a 1:100 dilution of the secondary mouse anti-FITC antibody (clone FL-D6, Cat# A1812, Sigma). The HiDefTM HRP-polymer system was used for detection (Cell Marque, Rocklin, CA). Endogenous peroxidase activity was eliminated with treatment under mild conditions as follows: 1.8% H2O2 for 5 min, 1% periodate for 5 min, and 0.02% NaBH4 for 2 min (Polak and Van Noorden, 2003 ). Signal detection was based on the Dako DAB chromogen kit according to the manufacturer's guidelines. We found that hot citric acid and slow cooling of the slide/citrate solution to room temperature (> 20 min) was the best method to retrieve the epitope, but lost epitope if we cooled quickly. The complete methodology can be found in Supplemental Experimental Procedure 1.
Clone fl d6
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The Clone FL-D6 is a specialized laboratory instrument designed for reliable and consistent DNA cloning and amplification. It is a core component in molecular biology workflows, enabling the replication of specific DNA sequences. The Clone FL-D6 performs its core function through precise temperature control and automated cycling processes.
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2 protocols using clone fl d6
1
Detecting Human Glioblastoma Cells in Mouse Brain
2
Phalloidin Staining and Electron Microscopy of Rat Spinal Cord
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Antibodies and Animals. FITC-binding phalloidin (P5282) and biotin-conjugated antibody against FITC (Clone FL-d6; Sigma) were obtained from Sigma (St. Louis, MO). This study used 5 adult SD rats (female 380 -420 g), which was approved by the Animal Research Committee of Jilin University, and handled in accordance with the NIH Guide for the Care and Use of Laboratory Animals.
Phalloidin staining and electron microscopy. The animals were perfused, and processed as described previously by us (Li et al., 2009) . Spinal cords, DRGs, and spinal nerves were dissected out at the level of L4 ~ L5, and cut into sections about 50 mm thick by a vibratome. The sections were then stained with FITC-conjugated phalloidin, which were further converted to peroxidase/DAB products by a FITC-anti-FITC system (Li et al., 2009) .
Light and ultrastructural observations. The phalloidin stained sections were examined with a laser scanning confocal microscope (Olympus FV1000), as described previously by us (Li et al., 2009) . The DAB stained sections were photographed using an Olympus BX53F microscope equipped with a digital imaging system (Olympus, Tokyo, Japan). Ultrathin sections were examined and photographed with a transmission electron microscope (JEOL JEM 1200EXII).
Phalloidin staining and electron microscopy. The animals were perfused, and processed as described previously by us (Li et al., 2009) . Spinal cords, DRGs, and spinal nerves were dissected out at the level of L4 ~ L5, and cut into sections about 50 mm thick by a vibratome. The sections were then stained with FITC-conjugated phalloidin, which were further converted to peroxidase/DAB products by a FITC-anti-FITC system (Li et al., 2009) .
Light and ultrastructural observations. The phalloidin stained sections were examined with a laser scanning confocal microscope (Olympus FV1000), as described previously by us (Li et al., 2009) . The DAB stained sections were photographed using an Olympus BX53F microscope equipped with a digital imaging system (Olympus, Tokyo, Japan). Ultrathin sections were examined and photographed with a transmission electron microscope (JEOL JEM 1200EXII).
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