Diets containing OVA were generated by modifying the powdered AIN-76 diet (American Institute of Nutrition, 1977) . Either 5% (wt/wt) casein was replaced with OVA (Wako Pure Chemical Industry, Osaka, Japan) as a protein source (OVA diet), or 6% (wt/ wt) casein was replaced with 5% (wt/wt) OVA and 1% (wt/wt) MBP (MBP/OVA diet). Seven-to nineweek-old male BALB/cA mice and OVA23-3 mice were separated into 3 groups: BALB/cA mice fed the OVA diet (OVA/BALB group), and OVA23-3 mice fed the OVA diet (OVA/OVA23-3 group) or the MBP/OVA diet (MBP/OVA/OVA23-3 group). The mice were housed individually and given free access to the diets throughout the experimental period. Body weights and food consumption were measured every 1 to 3 d. Blood was taken from the ocular fundus at d 10 and 28. At the end of the experimental period, the mice were killed by cervical dislocation and their lymphoid tissues [MLN, spleen, and bone marrow (BM)] and bones were removed for further analysis.
Ovalbumin (ova)
Manufactured by Fujifilm
Sourced in Japan
The OVA is a piece of laboratory equipment designed for the imaging and analysis of oocytes (eggs). It provides high-resolution imaging capabilities to support various research and clinical applications.
Automatically generated - may contain errors
Lab products found in correlation
Ovalbumin (ova), by Merck Group (1 mentions)
Isoflurane, by Fujifilm (1 mentions)
Ova grade 5, by Merck Group (1 mentions)
Balb c mice, by Hamamatsu Photonics (1 mentions)
May grunwald staining solution, by Fujifilm (1 mentions)
Isoflurane inhalation, by Fujifilm (1 mentions)
Aluminum hydroxide gel, by Fujifilm (1 mentions)
Superdex 200 increase 10 300 column, by GE Healthcare (1 mentions)
Kta explorer 10, by GE Healthcare (1 mentions)
Bovine serum albumin (bsa), by GE Healthcare (1 mentions)
Ferritin, by GE Healthcare (1 mentions)
Blue dextran, by GE Healthcare (1 mentions)
Aldolase, by GE Healthcare (1 mentions)
Hydrochloric acid (hcl), by Fujifilm (1 mentions)
Sodium hydroxide (naoh), by Fujifilm (1 mentions)
Bovine serum albumin (bsa), by Fujifilm (1 mentions)
Glycine, by Fujifilm (1 mentions)
Coelenterazine, by Fujifilm (1 mentions)
Chlorpromazine hydrochloride, by Fujifilm (1 mentions)
Bistris, by Dojindo Laboratories (1 mentions)
8 protocols using ovalbumin (ova)
1
Dietary OVA and MBP Effects on BALB/cA and OVA23-3 Mice
2
Murine Model of Allergic Sensitization
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Sensitization, SCIT, and challenges were conducted in accordance with our previous report [37 (link)], as follows (Figure 5 ). Briefly, 5-week-old BALB/c mice (Japan SLC, Hamamatsu, Shizuoka, Japan) were sensitized by i.p. injections with OVA (Grade V; Sigma-Aldrich, St. Louis, MO, USA) adsorbed to Al(OH)3 at a dose of 50 μg OVA/2 mg Al(OH)3/0.5 mL saline/animal on days 0 and 14. Seven days after the second sensitization, 0.5% OVA solution was injected subcutaneously at 200 μL (1 mg OVA)/animal on days 21, 23, and 25. The mice were intratracheally challenged with 0.02% OVA solution at 25 μL (5 μg OVA) on days 35, 36, 37, and 40 under inhalation anesthesia with isoflurane (Fujifilm, Osaka, Japan).
3
Allergic Sneeze Induction Protocol
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Female wilt-type BALB/c mice (8 week old) and male Hartley guinea pigs weighing 330–350 g were used. Mice were sensitized by an intraperitoneal injection of 2 mg aluminum hydroxide hydrate (ALUM; Cosmo Bio Co., Tokyo, Japan) and 10 μg ovalbumin (WAKO, Osaka, Japan) on days 0, 7, and 14. Guinea pigs were sensitized by 100 mg ALUM and 10 μg ovalbumin on day 0. Sneeze was induced by intranasal instillation of ovalbumin (100 μg/10 μl for mice, 150 μg/15 μl for guinea pigs) on days 21–28 [13 (link), 14 (link)].
4
Ovalbumin-based Allergic Asthma Model
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The following reagents were purchased: phosphate buffered saline (PBS) solution (Nissui Pharmaceutical Co., Tokyo, Japan); ovalbumin (OA), aluminum hydroxide gel, isoflurane inhalation, May-Grunwald staining solution, and Giemsa’s staining solution (Wako Pure Chemical Industries, Osaka, Japan).
5
Size Exclusion HPLC of Recombinant Peptidases
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Recombinant peptidases were subjected to size exclusion HPLC using ÄKTA explorer 10S (GE Healthcare, Chicago, U.S.A.) with a Superdex200 increase 10/300 column (1.5 X 30 cm) equilibrated with 20 mM Tris-HCl (pH 8.0) containing 0.1 M NaCl. They were eluted with the identical buffer at a rate of 0.5 ml/min at room temperature, then 0.5-ml fractions were collected. An aliquot (1 µl) of the fractions was subjected to a peptidase assay with Z-Lys-Met-MCA. Blue dextran (void volume), ferritin (430 kDa), aldolase (182 kDa), bovine serum albumin (67 kDa), (GE Healthcare), ovalbumin (45 kDa) (Wako Pure Chemicals, Osaka, Japan), and cytochrome C (12 kDa) (Sigma-Aldrich) were used as molecular standards.
6
Bioluminescent Protein Assay Reagents
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The commercially available materials used in this study were obtained from the following commercial suppliers. Human alpha 1-acid glycoprotein (alpha 1-acid glycoprotein from human plasma; lot number: SLBJ6840V) was from Sigma-Aldrich (St. Louis, MO, USA). OVA (albumin from egg; lot number: CAL1231), BSA (albumin from bovine serum, fatty acid/IgG/protease free; lot number: CAQ4454), coelenterazine, chlorpromazine hydrochloride, Tris-HCl buffers, glycine, sodium hydroxide, and hydrochloric acid were from FUJIFILM Wako Pure Chemical Corporation (Osaka, Japan). HSA (albumin, human serum, F-V; lot number: M8H7013) was from Nacalai Tesque (Kyoto, Japan). Cypridina luciferin was from ATTO Corporation (Tokyo, Japan). BisTris was from Dojindo Laboratories. All materials were used without further purification. A recombinant CLase from C. noctiluca was prepared according to the method reported previously [12 (link)].
7
PCA Reaction Induction and Quantification
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The PCA reaction model was prepared as described previously [12 (link)]. In brief, OVA (Seikagaku Corporation, Tokyo, Japan) was suspended in 20 mg/ml Al(OH)3 solution (final concentration of OVA, 10 μg/ml), and 0.2 ml of this suspension (OVA, 2 μg) was injected intraperitoneally into 4-week-old female Balb/c mice five times at 2-week intervals. 10 days after the fifth injection of OVA, whole blood was collected to obtain anti-OVA serum. A 10-μl aliquot of 30-fold diluted anti-OVA serum was injected intracutaneously into each ear of 6-week-old male ddY mice After 48 h, OVA (10 mg/kg body weight) in saline containing 0.5 % Evans blue (Wako Pure Chemicals) was injected intravenously to induce PCA reaction (antigen challenge). The BKT samples were suspended in water and administered orally to overnight-fasted mice 2 h before antigen challenge. The mice were killed by cervical dislocation 30 min after the antigen challenge. Their ears were removed, immersed in 0.3 ml of 1 M KOH overnight at 37 °C, and Evans blue in the ears was extracted with 0.7 ml of acetone/3.3 M phosphoric acid (67:3). Evans blue concentration was measured colorimetrically at 620 nm. Data are expressed as quantity of Evans blue per gram fresh weight of ears.
8
FITC-dextran and FITC-ovalbumin Conjugation
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Fluorescein isothiocyanate (FITC)-dextran (FD4, MW 4,000) and FITC-OVAlbumin conjugate (FITC-OVA, MW45,000) were obtained from Sigma-Aldrich Japan (Tokyo, Japan) and Thermo Fisher Scientific (Tokyo, Japan), respectively. OVA was obtained from FUJIFILM Wako Pure Chemicals (Osaka, Japan). Sodium dodecyl sulfate (SDS) was obtained from Kanto Chemicals (Tokyo, Japan).
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