Erythromycin (ERY), erythromycin ethyl succinate (ESE), clarithromycin (CLA), roxithromycin (ROX), azithromycin (AZI), tulathromycin (TUL), oleandomycin (OLE), 1,2-ethylenediamine (EDA), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), N,N′-dicyclohexylcarbodiimide (DCC), N-hydroxysuccinimide (NHS), carboxymethoxylamine hemihydrochloride (CMO), sodium periodate (PI), sodium borohydride, dimethyl adipimidate (DMA), bovine serum albumin (BSA), gelatin (GEL), and horseradish peroxidase (HRP) were purchased from Chimmed (Moscow, Russia). Goat anti-rabbit IgG antibodies conjugated to horseradish peroxidase (GAR-HRP) were purchased from Imtek Ltd. (Moscow, Russia). Dimethylformamide (DMF), glutaraldehyde (GA), and ovalbumin (OVA) were obtained from Serva (Heidelberg, Germany). 9-(Carboxymethyloxime)-clarithromycin (cmoCLA) and conjugated antigens and antibodies against BSA-cmoCLA(ae) were prepared and described in our previous work [18 (link)].
Ovalbumin (ova)
Manufactured by Serva Electrophoresis
Sourced in Germany
The OVA is a compact and efficient electrophoresis system designed for the analysis of nucleic acids and proteins. It features a simple and user-friendly interface, enabling researchers to perform reliable and reproducible separations. The OVA is a versatile laboratory tool that supports a range of applications in molecular biology, genetics, and biochemistry.
Automatically generated - may contain errors
Lab products found in correlation
Bovine serum albumin (bsa), by Merck Group (2 mentions)
Tween 20, by Merck Group (2 mentions)
O phenylenediamine dihydrochloride, by Merck Group (2 mentions)
Monoclonal anti chicken egg albumin clone ova 14 mouse igg1 isotype, by Merck Group (2 mentions)
Muramyl dipeptide mdp, by InvivoGen (1 mentions)
Ovalbumin (ova), by Worthington (1 mentions)
Aluminium hydroxide, by Serva Electrophoresis (1 mentions)
Ova grade 5, by Merck Group (1 mentions)
Imject alum, by Thermo Fisher Scientific (1 mentions)
Citric acid monohydrate, by Merck Group (1 mentions)
Pyrilamine maleate, by Merck Group (1 mentions)
Olodaterol hydrochloride, by Boehringer Ingelheim (1 mentions)
Formoterol fumarate, by Boehringer Ingelheim (1 mentions)
Dicetyl phosphate, by Merck Group (1 mentions)
8 protocols using ovalbumin (ova)
1
Antibody Production and Characterization
2
NOD2 Agonist Synthesis and Characterization
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Muramyl dipeptide (MDP) was obtained from Invivogen, Inc., (San Diego, CA, USA). The lipidated NOD2 agonists (SG29, SG115, and ZSB63) were synthesized as described [8 (link),24 (link)]. The HPLC purity of all pharmacologically investigated compounds was >95%. Bovine serum albumin, Tween 20, monoclonal anti-chicken egg albumin (clone OVA-14 mouse IgG1 isotype), and o-phenylenediaminedihydrochloride were obtained from Sigma (St. Louis, MO, USA). Horseradish-peroxidase-conjugated goat anti-mouse IgG (HRP-anti-mouse IgG) was obtained from Bio-Rad Laboratories (Hercules, CA, USA). Monoclonal antibodies (biotin-conjugated rat anti-mouse IgG1 and anti-mouse IgG2a) and streptavidin-HRP conjugate were purchased from PharMingen, Becton Dickinson (Franklin Lakes, NJ, USA). Chemicals used for the preparation of solutions and buffers were purchased from Kemika (Zagreb, Croatia). Ovalbumin (OVA) was purchased from Serva (Heidelberg, Germany). Egg-phosphatidylcholine (L-α-Phosphatidylcholine, type XI-E) was purchased from Avanti Polar Lipids. Cholesterol from the porcine liver was purchased from Sigma (USA).
3
Murine Model of Allergic Sensitization
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Twenty BALB/c mice were divided into four groups and treated with PBS, OVA, OVA + L900/2 or OVA + L900/3 (4–6 mice per group). Mice were sensitized by intraperitoneal (i.p.) injections of 10 mg of OVA (Worthington, Lakewood, NJ, USA) or 10 mg of OVA + 60 μg of each polysaccharide adsorbed to 100 μl of aluminium hydroxide (Alum, Serva, Germany) in a final volume of 200 μl, on days 1 and 14. Control mice received 100 μl PBS and 100 μl aluminium hydroxide. On day 7 after the last immunization, the mice were euthanized by cervical dislocation. Blood was collected, and serum samples were stored at −40°C until analysis. Mesenteric lymph nodes (MLN, pooled per group) and spleens were aseptically removed and prepared for cytokine determination. All experiments were repeated twice using four to six mice per group each time.
4
OVA Immunization Protocol for Mice
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Sex-matched NIH/OlaHsd mice (five
per group) were immunized subcutaneously into the tail base with OVA
(10 μg; Serva, Germany) alone or plus the compounds (50 μg)
on days 0, 21, and 42. The injection volume in all of the experimental
groups was 0.1 mL per mouse. On the seventh day after the second booster
dose, the mice were anesthetized with i.p. application of ketamine/xylazine
(25 mg/kg each) prior to blood collection from the axillary plexus.
Individual sera from each animal were decomplemented at 56 °C
for 35 min and then stored at −20 °C until tested.
per group) were immunized subcutaneously into the tail base with OVA
(10 μg; Serva, Germany) alone or plus the compounds (50 μg)
on days 0, 21, and 42. The injection volume in all of the experimental
groups was 0.1 mL per mouse. On the seventh day after the second booster
dose, the mice were anesthetized with i.p. application of ketamine/xylazine
(25 mg/kg each) prior to blood collection from the axillary plexus.
Individual sera from each animal were decomplemented at 56 °C
for 35 min and then stored at −20 °C until tested.
5
Induction and Analysis of Murine Allergic Airway Disease
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As previously published for development of EAAD [7, 9] , mice (B6.SJL-Ptprca as donors for chimeric mice or fully engrafted chimeric mice) were injected intraperitoneally with OVAlbumin (OVA) (Sigma-Aldrich, St Louis, MO) in alum (aluminium hydroxide suspension, Serva, Heidelberg, Germany) on day 0 (10 µg OVA with 2 mg alum in 200 µl saline per mouse), and again on day 14. These mice were then challenged for 30 min on day 21 with a 1% OVA-in-saline aerosol (UltraNeb, DeVibiss, Somerset, PA). Twenty-four hours later, some mice were euthanased, and the BM harvested for transfer to new mice. Alternatively, mice were further challenged two times with OVA aerosol on days 22 and 23 and 24 hours after the last challenge, the mice were euthanased, and airway draining lymph nodes (ADLN; mediastinal, tracheobrachial, and parathymic) and bronchoalveolar lavage fluid (BALF) collected.
6
Ovalbumin-Induced Pulmonary Th2 Inflammation
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After systemic sensitization via i.p. injections of 20 μg of ovalbumin (OVA; grade V; Merck KGaA) dissolved in 100 µL of aluminum hydroxide suspension (1.3%; SERVA Electrophoresis GmbH, Heidelberg, Germany) and 10 µL of phosphate-buffered saline (PBS) on days 0 and 14, mice were repeatedly exposed to aerosolized ovalbumin (1%) in PBS on days 28–30 for 20 min/day (53 (link), 54 (link)). On the respective days, sham-treated mice received i.p. injections of 100 µL of aluminum hydroxide suspension and 10 µL of PBS, followed by exposure to aerosolized PBS. Effects of pulmonary Th2 inflammation were analyzed on day 32.
7
Ovalbumin-Induced Airway Inflammation Protocol
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Ovalbumin (Serva Electrophoresis GmbH, Heidelberg, Germany) was dissolved and diluted in phosphate buffered saline (PBS; Lonza, Verviers, Belgium) for the immunization, and in 0.9% saline (Delta Select GmbH, Dreieich, Germany) for the OVA challenge. Aluminium hydroxide (ImjectAlum; ThermoScientific, Rockford, IL, USA) was dissolved and diluted in PBS. Citric acid monohydrate and pyrilamine maleate were purchased from Sigma-Aldrich Chemie GmbH (Steinheim, Germany) and dissolved and diluted in 0.9% saline. Respitose (Lactose monohydrate, ML003) was from Boehringer Ingelheim (Ingelheim, Germany). Codeine phosphate hemihydrate was purchased from Merck Company (Germany). Olodaterol hydrochloride, salmeterol xinafoate, formoterol fumarate, and indacaterol maleate were synthesized at the Department of Chemical Research, Boehringer Ingelheim Pharma GmbH & Co. KG (Biberach, Germany).
8
Immunoassay Reagent Preparation Protocol
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Bovine serum albumin, Tween 20, monoclonal
anti-chicken egg albumin
(clone OVA-14 mouse IgG1 isotype), o-phenylenediaminedihydrochloride,
and MDP were from Sigma (U.S.A.). Horseradish-peroxidase-conjugated
goat anti-mouse IgG (HRP-anti-mouse IgG) was from Bio-Rad Laboratories
(U.S.A.). Biotin-conjugated rat anti-mouse IgG1 and anti-mouse IgG2a
monoclonal antibodies and streptavidin–peroxidase were from
PharMingen, Becton Dickinson (U.S.A.). Chemicals for buffers and solutions
were from Kemika (Croatia). Ovalbumin was from Serva (Germany).l -α-Phosphatidylcholine, type XI-E, from fresh egg yolk
(egg-phosphatidylcholine) was from Avanti Polar Lipids. Cholesterol
from porcine liver and dicetylphosphate were from Sigma (U.S.A.).
Monomannosyl-PEG-palmitic acid derivative (Man-PEG-Pam) was synthesized
as previously described.129 (link)
anti-chicken egg albumin
(clone OVA-14 mouse IgG1 isotype), o-phenylenediaminedihydrochloride,
and MDP were from Sigma (U.S.A.). Horseradish-peroxidase-conjugated
goat anti-mouse IgG (HRP-anti-mouse IgG) was from Bio-Rad Laboratories
(U.S.A.). Biotin-conjugated rat anti-mouse IgG1 and anti-mouse IgG2a
monoclonal antibodies and streptavidin–peroxidase were from
PharMingen, Becton Dickinson (U.S.A.). Chemicals for buffers and solutions
were from Kemika (Croatia). Ovalbumin was from Serva (Germany).
(egg-phosphatidylcholine) was from Avanti Polar Lipids. Cholesterol
from porcine liver and dicetylphosphate were from Sigma (U.S.A.).
Monomannosyl-PEG-palmitic acid derivative (Man-PEG-Pam) was synthesized
as previously described.129 (link)
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