To analyse the expression of NLRP3 inflammasome, primary PMs were incubated for 3 h after seeding in the culture plate. Macrophages were assayed in opti‐MEM media (Gibco, 31985) in the presence or the absence of trehalose or saccharin 15 min prior to LPS stimulation (Invivogen, tlrl‐eklps; 10 ng/ml) for 3 h followed by adding ATP (Sigma–Aldrich, A6419; 5 mM) for 30 min to release IL‐1β. To confirm Atg7 protein deficiency in macrophage of Atg7fl/fl or Atg7fl/fl;Lyz2‐Cre mice, PMs were harvested, seeded, incubated for 3 h, and the adherent macrophages were collected for further analysis. Human T1R3 was analysed from cell lysates obtained from THP‐1 cell line or PBMCs, as described.86 Proteins in the culture supernatant were concentrated by chloroform‐methanol precipitation, as described previously.90 Primary antibodies used were goat anti‐mouse IL‐1β antibody (R&D Systems, AF‐401‐NA; 1:1000), mouse monoclonal anti‐NLRP3 antibody (Adipogen, AG‐20B‐0014; 1:1000), mouse monoclonal anti‐caspase‐1 antibody (AdipoGen, AG‐20B‐0042; 1:1000), rabbit anti‐Atg7 antibody (Cell Signaling, 2631; 1:1000), rabbit anti‐human T1R3 (abcam, ab229015; 1:1000), and mouse anti‐β‐Actin antibody (BD Biosciences, 612657; 1:5000).
Mouse anti
Manufactured by BD
Sourced in Belgium
Mouse anti is a laboratory reagent used for the detection and identification of specific target molecules or antigens in biological samples. It functions as a primary antibody, binding to the target antigen and enabling its subsequent detection or isolation.
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Lab products found in correlation
2 protocols using mouse anti
1
Analyzing NLRP3 Inflammasome in Macrophages
2
Antibody Reagents for SHIP2 Regulation Studies
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Anti-SKIP rabbit polyclonal antibody was from LifeSpan BioSciences, Inc (BioConnect, The Netherlands; cat.no. LS-B10239). SHIP2 antibodies used in Western blot were from CST (cat. no. 2839) or made against a C-terminal peptide of human SHIP2 (Muraille et al., 1999) . For SHIP2 immunostaining, we used either the CST rabbit antibody or the Novus mouse monoclonal (cat.no. H00003636-M01). Anti-PI(4,5)P2 (2C11) (cat.no. Z-P045) antibody was from Echelon Biosciences, Inc (Le-Perray-en-Yvelines, France). Mouse anti- ) was from BD Biosciences (Erembodegem, Belgium). Lipofectamine RNAi Max (cat.no. 13778030), calnexin monoclonal antibody (cat.no. MA3-027-A488), normal horse serum (NHS) (cat.no. 26050-070), Alexa-Fluor-488-conjugated donkey (cat.no. A21202), Alexa-Fluor-594-conjugated goat (cat.no. A11012) anti-rabbit-IgG, Alexa-Fluor-488-conjugated donkey (cat.no. A21206) and Alexa-Fluor-594-conjugated goat (cat.no. A11032) anti-mouse-IgG, Hoechst 33342 (cat.no. H-3570) were from Invitrogen (Breda, The Netherlands). The Glycergel mounting medium (cat.no. C0563) was from Dako (Heverlee, Belgium). Growth factor-reduced extracellular matrix (cat.no. E1270) and fibronectin from bovine plasma (cat.no. F1141) were from Sigma-Aldrich (Diegem, Belgium). SMARTpool Silencer siRNA against human IPMK (cat.no. L-006740-00-0005) and Non-targeting siRNA (cat.no. D-001810-01-05) were from GE/Dharmacon.
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