Anti-inflammatory Macrophage Activation Protocol

Schneider’s insect medium, lipopolysaccharide (LPS), IFN-γ, resazurin sodium salt, the ornithine decarboxylase (ODC) antibody and BHT (butylated hydroxytoluene) were all obtained from SIGMA-Aldrich (St Louis, MO, United States). Lactate Dehydrogenase (LDH) Cytotoxicity Detection Kit and Nutridoma™-SP supplement were obtained from Roche Diagnostics GmbH (Sandhofer Strasse, Mannheim, Germany). Inactive fetal bovine serum (FBS), RPMI 1640 medium and RPMI 1640 medium without phenol red and penicillin were purchased from GIBCO (Carlsbad, CA, United States). Streptomycin, L-glutamine and PBS 10x were obtained from Invitrogen (Carlsbad, CA, United States). The superoxide dismutase activity assay, NS398 COX-2 inhibitor and PGE₂ Elisa kits were purchased from Cayman Chemical Company (Ann Arbor, Michigan, United States). The MILLIPLEX^® kit based on Luminex^® xMAP^® technology was obtained from Merck (Darmstadt, Germany). A protease and phosphatase inhibitor cocktail, T-PERTM Tissue Protein Extraction Reagent and SuperSignal™ West Atto Ultimate Sensitivity Substrate Kit were purchased from ThermoFisher Scientific (Waltham, MA United States). Primary β-actin, pERK and ERK antibodies were obtained from Cell Signaling (Danvers, MA, United States), while Cox-2 and iNOS antibodies were purchased from Calbiochem (San Diego, CA, United States). The HO-1 antibody was purchased from Enzo Life Science (Farmingdale, NY, United States). Arginase antibody was obtained from R&D Systems (Minneapolis, MN, United States). Nrf2 antibody was obtained from Abcam (Cambridge, United Kingdom). Propylene glycol was purchased from Rudnik Comércio de Produtos Químicos Ltda. (Cotia, SP, BR). Carbomer 940 (Carbopol^®) and sodium metabisulphite were obtained from Henrifarma Produtos Químicos (São Paulo, SP, BR). Potassium sorbate was purchased from Cosmoquímica Indl. E Coml. Ltda. (São Paulo, SP, BR). Disodium EDTA was obtained from Zílquimica Produtos para Laboratórios Ltda. (Ribeirão Preto, SP, BR). Castor oil was purchased from Volp Indústria e Comércio Ltda. (Osasco, SP, BR). Triethanolamine was obtained from Química Moderna Indústria e Comércio Eireli (Barueri, SP, BR).

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Rebouças-Silva J., Amorim N.A., Jesus-Santos F.H., de Lima J.A., Lima J.B., Berretta A.A, & Borges V.M. (2023). Leishmanicidal and immunomodulatory properties of Brazilian green propolis extract (EPP-AF®) and a gel formulation in a pre-clinical model. Frontiers in Pharmacology, 14, 1013376.

Publication 2023

Actin Antibodies Antibody Arginase Assay Butylated hydroxytoluene Carbomer 940 Carbopol Castor oil Cayman Cox 2 Cox 2 inhibitor Cytotoxicity Diagnostics Disodium edta Elisa Fetal bovine serum Ifn γ Inos Insect L glutamine Lactate dehydrogenase Lipopolysaccharide Nrf2 Ns398 Ornithine decarboxylase Penicillin Pge2 Phosphatase Potassium sorbate Propylene glycol Protease Resazurin Salt Sensitivity Sodium Sodium metabisulphite Streptomycin Superoxide dismutase Supplement T protein Tissue Triethanolamine

Corresponding Organization :

Other organizations : Universidade do Estado da Bahia, Universidade Federal do Oeste da Bahia

Top 5 similar protocols

Variable analysis

independent variables

Schneider's insect medium
Lipopolysaccharide (LPS)
IFN-γ
NS398 COX-2 inhibitor

dependent variables

Resazurin sodium salt
Lactate Dehydrogenase (LDH) Cytotoxicity
Superoxide dismutase activity
PGE2 levels
Protein expression (β-actin, pERK, ERK, Cox-2, iNOS, HO-1, Arginase, Nrf2)

control variables

Inactive fetal bovine serum (FBS)
RPMI 1640 medium
RPMI 1640 medium without phenol red and penicillin
Streptomycin
L-glutamine
PBS 10x
Nutridoma™-SP supplement
Protease and phosphatase inhibitor cocktail
T-PERTM Tissue Protein Extraction Reagent
SuperSignal™ West Atto Ultimate Sensitivity Substrate Kit
Propylene glycol
Carbomer 940 (Carbopol®)
Sodium metabisulphite
Potassium sorbate
Disodium EDTA
Castor oil
Triethanolamine

positive controls

BHT (butylated hydroxytoluene)

negative controls

Not specified

Annotations

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