OV2944 cells were treated with neuraminidase (1 μM, 37 °C, 1 h, Fujifilm Wako, Japan) to eliminate terminal sialic acid residue of GM1b. Cells were treated with biotin-labeled Maackia amurensis II (MLAII, 50 μg/ml, vector labs, FITC-labeled wheat germ agglutinin (WGA, 50 μg/ml, vector labs, 30 min, RT). MLAII recognized sialic acid, whereas WGA did not. After fixation, cells were treated with Texas-red streptavidin (1 μg/ml, 30 min, RT). Following fixation, cell were treated with rhodamine-labeled p6 (Rhod-P6) and anti-CD44 antibody (Abcam, ab25340, 1:100), and reacted with secondary antibody and mounted.
Ab25340
Manufactured by Abcam
Sourced in United States
Ab25340 is a laboratory antibody product offered by Abcam. It is a polyclonal antibody raised in Rabbit. The product is designed for use in various immunoassay techniques.
Automatically generated - may contain errors
3 protocols using ab25340
1
Sialic Acid Residue Visualization
2
Fluorescent Antibody Staining for LYVE-1
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Primary antibodies used were FITC-conjugated rabbit polyclonal anti-group A carbohydrate (Abcam ab68879. 0.2 mg/ml), goat polyclonal anti-human LYVE-1 (R&D AF2089), and goat polyclonal anti-mouse LYVE-1 (R&D AF2125). Blocking antibodies mAb20891 (R&D) and (BRIC235 IBGRL) were used against human LYVE-1 and CD44 respectively and mAb2125 (R&D) and ab25340 (KM201, Abcam) were used against mouse LYVE-1 and CD44 respectively. Blocking antibodies and isotype controls were used at 20 μg/ml.
3
Mandibular Bone Healing Assessment
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Three and seven days after the application on the socket, the animals were killed and the mandibular preparation was taken and fed into a 10% formaldehyde buffer solution to prevent the tissue from decomposing, tissue hardening, increasing the refractive index of various tissue components, and increasing the affinity of the tissue against the stain. After the process of tissue fixation, the process of decalcification was done using EDTA for 1 month. Mandibular specimens were prepared in the form of transversal preparations with hematoxylin-eosin and immunohistochemical staining with monoclonal anti-CD44 (ab25340, Abcam, United States) and monoclonal anti-IL-10 (ab189392, Abcam). After that, we observed the number of blood vessels and the number of macrophage cells reacted positively to anti-CD44 and anti-IL-10 in the socket with a light microscope (Olympus CX21, Japan) at 100 and 400 magnification. Furthermore, data tabulation and statistical analysis were conducted with one-way analysis of variance followed by the Tukey’s honestly significant difference (HSD) test.
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