Wiener reagents and procedures

Manufactured by Wiener Lab

Sourced in Argentina

Wiener reagents and procedures are a set of laboratory products designed for use in various analytical and testing applications. The core function of these reagents and procedures is to facilitate the accurate and precise measurement and analysis of samples. The specific details and intended uses of these products are not included in this factual and unbiased description.

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Lab products found in correlation

Albumin diagnostic kit, by Wiener Lab (5 mentions) Elisa technique kits, by R&D Systems (1 mentions) Cell proliferation kit 1, by Roche (1 mentions)

8 protocols using wiener reagents and procedures

Pulmonary Barrier Permeability Assessment

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Broncho-alveolar lavages (BAL) samples were obtained as described previously (16 (link), 17 (link)). Briefly, the trachea was exposed and intubated with a catheter, and 2 sequential lavages were performed in each mouse by injecting sterile PBS. The recovered fluid was centrifuged for 10 min at 900 g; and frozen at −70°C for subsequent analyses.
Albumin content, a measure to quantitate increased permeability of the bronchoalveolar–capillarity barrier, and lactate dehydrogenase (LDH) activity, an indicator of general cytotoxicity, were determined in the acellular BAL fluid. Albumin content was determined colorimetrically based on albumin binding to bromcresol green using an albumin diagnostic kit (Wiener Lab, Buenos Aires, Argentina). LDH activity, expressed as units per liter of BAL fluid, was determined by measuring the formation of the reduced form of nicotinamide adenine dinucleotide (NAD) using the Wiener reagents and procedures (Wiener Lab).

Garcia-Castillo V., Tomokiyo M., Raya Tonetti F., Islam M.A., Takahashi H., Kitazawa H, & Villena J. (2020). Alveolar Macrophages Are Key Players in the Modulation of the Respiratory Antiviral Immunity Induced by Orally Administered Lacticaseibacillus rhamnosus CRL1505. Frontiers in Immunology, 11, 568636.

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Evaluating Lung Barrier Integrity via BAL

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Broncho-alveolar lavages (BAL) samples were obtained as described previously [17 (link),22 (link)]. Briefly, the trachea was exposed and intubated with a catheter, and 2 sequential lavages were performed in each mouse by injecting sterile PBS. The recovered fluid was centrifuged for 10 min at 900× g and frozen at −70 °C for subsequent analyses.
Albumin content, a measure to quantitate the increased permeability of the bronchoalveolar–capillarity barrier, and lactate dehydrogenase (LDH) activity, an indicator of general cytotoxicity, were determined in the acellular BAL fluid. Albumin content was determined colorimetrically based on albumin binding to bromcresol green using an Albumin Diagnostic Kit (Wiener Lab, Buenos Aires, Argentina). LDH activity, expressed as units per liter of BAL fluid, was determined by measuring the formation of the reduced form of nicotinamide adenine dinucleotide (NAD) using the Wiener reagents and procedures (Wiener Lab).

Clua P., Tomokiyo M., Raya Tonetti F., Islam M.A., García Castillo V., Marcial G., Salva S., Alvarez S., Takahashi H., Kurata S., Kitazawa H, & Villena J. (2020). The Role of Alveolar Macrophages in the Improved Protection against Respiratory Syncytial Virus and Pneumococcal Superinfection Induced by the Peptidoglycan of Lactobacillus rhamnosus CRL1505. Cells, 9(7), 1653.

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Evaluation of Pulmonary Tissue Injuries

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For the evaluation of pulmonary tissue injuries, lungs were excised, washed out with PBS, and immersed in 4% (v/v) formalin saline solution for the histologycal study as previously mentioned [15 (link)]. In addition, biochemical parameters as albumin content and lactate dehydrogenase (LDH) activity were studied in broncho-alveolar lavages (BAL). Albumin content was determined colorimetrically based on albumin binding to bromocresol green using an albumin diagnostic kit (Wiener Lab). LDH activity was determined by measuring the formation of the reduced form of nicotinamide adenine dinucleotide using the Wiener reagents and procedures (Wiener Lab), and was expressed as units per liter of BAL fluid [18 (link)].

Kolling Y., Salva S., Villena J, & Alvarez S. (2018). Are the immunomodulatory properties of Lactobacillus rhamnosus CRL1505 peptidoglycan common for all Lactobacilli during respiratory infection in malnourished mice?. PLoS ONE, 13(3), e0194034.

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Quantifying Alveolar Barrier Permeability

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Albumin content, a measure to quantitate increased permeability of the bronchoalveolar–capillarity barrier, and lactate dehydrogenase (LDH) activity, an indicator of general cytotoxicity, were determined in BAL of infected animals as described previously [16 (link), 17 (link)]. Briefly, albumin content was determined colorimetrically based on albumin binding to bromcresol green using an albumin diagnostic kit (Wiener Lab, Buenos Aires, Argentina). LDH activity, expressed as units per liter of BAL fluid, was determined by measuring the formation of the reduced form of nicotinamide adenine dinucleotide (NAD) using the Wiener reagents and procedures (Wiener Lab).

Laiño J., Villena J., Suvorov A., Zelaya H., Ortiz Moyano R., Salva S, & Alvarez S. (2018). Nasal immunization with recombinant chimeric pneumococcal protein and cell wall from immunobiotic bacteria improve resistance of infant mice to Streptococcus pneumoniae infection. PLoS ONE, 13(11), e0206661.

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Evaluating Toxicity in Poly(I:C) Challenged Mice

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Lactate dehydrogenase (LDH) and aspartate aminotransferase (AST) activities were determined in the serum to evaluate general toxicity of poly(I:C) in mice challenged by the intraperitoneal injection. Blood samples were obtained through cardiac puncture under anesthesia. LDH and AST activities, expressed as units per liter of serum, were determined by measuring the formation of the reduced form of nicotinamide adenine dinucleotide (NAD) using the Wiener reagents and procedures (Wiener Lab, Buenos Aires, Argentina) (11 (link)).

Mizuno H., Arce L., Tomotsune K., Albarracin L., Funabashi R., Vera D., Islam M.A., Vizoso-Pinto M.G., Takahashi H., Sasaki Y., Kitazawa H, & Villena J. (2020). Lipoteichoic Acid Is Involved in the Ability of the Immunobiotic Strain Lactobacillus plantarum CRL1506 to Modulate the Intestinal Antiviral Innate Immunity Triggered by TLR3 Activation. Frontiers in Immunology, 11, 571.

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Evaluation of Gastrointestinal Injury

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Lactate dehydrogenase (LDH) and aspartate aminotransferase (AST) activities were determined in the serum to evaluate gastrointestinal injury indirectly. Blood samples were obtained through cardiac puncture under anesthesia. LDH and AST activities, expressed as units per liter of serum, were determined by measuring the formation of the reduced form of nicotinamide adenine dinucleotide (NAD) using the Wiener reagents and procedures (Wiener Lab, Buenos Aires, Argentina) (38 (link), 39 (link)).
Intestinal fluid samples were obtained as described before (38 (link), 39 (link)). Briefly, the small intestine was flushed with 5 ml of PBS and the fluid was centrifuged (10,000 g, 4°C 10 min) to separate particulate material. The intestinal supernatant samples were kept frozen at -80°C until use. Tumor necrosis factor (TNF)-α, IL-6, IL-10, IL-15, interferon (IFN)-β and IFN-γ, chemokine KC (or CXCL1), and MCP-1 concentrations in intestinal fluid a were measured with commercially available enzyme-linked immunosorbent assay (ELISA) technique kits following the manufacturer’s recommendations (R&D Systems, MN, USA).

Indo Y., Kitahara S., Tomokiyo M., Araki S., Islam M.A., Zhou B., Albarracin L., Miyazaki A., Ikeda-Ohtsubo W., Nochi T., Takenouchi T., Uenishi H., Aso H., Takahashi H., Kurata S., Villena J, & Kitazawa H. (2021). Ligilactobacillus salivarius Strains Isolated From the Porcine Gut Modulate Innate Immune Responses in Epithelial Cells and Improve Protection Against Intestinal Viral-Bacterial Superinfection. Frontiers in Immunology, 12, 652923.

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Cytotoxicity and Viability Assays

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The cytotoxic effect of the different treatments on AGS and THP-1 cells was evaluated by the determination of LDH in the culture medium using the Wiener reagents and procedures (Wiener Lab, Buenos Aires, Argentina).
In addition, cells viability was evaluated with the Bromide reduction assay of 3 (4,5-dimethyl-2-thiazoyl)-2,5diphenyltetrazole (MTT) Roche Cell Proliferation Kit I (Roche Diagnostics GmbH, Mannheim, Germany) following the manufacturer's instructions.

Garcia-Castillo V., Zelaya H., Ilabaca A., Espinoza-Monje M., Komatsu R., Albarracín L., Kitazawa H., Garcia-Cancino A, & Villena J. (2018). Lactobacillus fermentum UCO-979C beneficially modulates the innate immune response triggered by Helicobacter pylori infection in vitro. Beneficial microbes, 9(5).

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Comprehensive Lung Function Assessment

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Protein content was measured by the bicinchoninic protein assay (Pierce Biotechnology Inc., Rockford, IL, USA). Albumin content was determined colorimetrically based on albumin binding to bromocresol green using an albumin diagnostic kit (Wiener Lab, Buenos Aires, Argentina).
LDH activity, expressed as units per liter of BAL fluid, was determined by measuring the formation of the reduced form of nicotinamide adenine dinucleotide using the Wiener reagents and procedures (Wiener Lab). The lung wet:dry weight ratio was measured as previously obtained and described by Aeffner et al. [18] . Briefly, mice were euthanized and exsanguinated, and their lungs removed, weighed, and dried in an oven at 55 °C for 7 days. After drying, the lungs were weighed again. Wet:dry weight ratio was then calculated as an index of intrapulmonary fluid accumulation, without correction for blood content.
Whole-lung samples from all experimental groups were excised and washed out with PBS. Then, tissues were immersed in 4 % (v/v) formalin saline solution. Once fixed, samples were dehydrated and embedded in Histowax (Leica Microsystems Nussloch GmbH, Nussloch, Germany) at 56 °C. Finally, lungs were cut into 4 lm serial sections and stained with hematoxylin-eosin for light microscopy examination. All slides were coded and evaluated blindly.

Zelaya H., Tada A., Vizoso-Pinto M.G., Salva S., Kanmani P., Agüero G., Alvarez S., Kitazawa H, & Villena J. (2015). Nasal priming with immunobiotic Lactobacillus rhamnosus modulates inflammation-coagulation interactions and reduces influenza virus-associated pulmonary damage. Inflammation research : official journal of the European Histamine Research Society ... [et al.], 64(8).

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