Frozen bacterial cells were centrifuged after thawing, and the pellet was resuspended in PBST. After anesthetizing C3HeB/FeJ mice, 1×106 CFU/mouse of Ms_Vec or Ms_AcpM were inoculated intranasally. At the indicated times after infection, mice were euthanized and the lungs were collected to assess the bacterial burden. Lung tissues were homogenized using a tissue homogenizer (Omni International Inc., Warrenton, VA, USA) in PBST. Serial dilutions of the homogenates were planted in 7H10 agar plates, and colonies were counted after 3-4 days of incubation at 37°C.
Tissue homogenizer
Manufactured by Omni International
Sourced in United States
The Tissue Homogenizer is a laboratory instrument used to disrupt and homogenize solid tissue samples. It employs a high-speed motor and rotating blade to rapidly break down and mix tissue into a homogeneous suspension, facilitating further analysis or processing.
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Lab products found in correlation
Trizol, by Thermo Fisher Scientific (5 mentions)
Rneasy mini kit, by Qiagen (3 mentions)
T per, by Thermo Fisher Scientific (2 mentions)
Lightcycler 480 instrument system, by Roche (2 mentions)
Purelink rna mini kit, by Thermo Fisher Scientific (2 mentions)
Pgc 1α, by Abcam (2 mentions)
Hrp conjugated secondary antibody, by Cell Signaling Technology (2 mentions)
Pgc1β, by Abcam (2 mentions)
Westernbright ecl kit solution, by Advansta (2 mentions)
Gapdh, by Biosynth (2 mentions)
Trizol reagent, by Thermo Fisher Scientific (2 mentions)
Ficoll paque plus, by GE Healthcare (2 mentions)
Human recombinant colony stimulation factor 1, by R&D Systems (2 mentions)
Nanodrop 2000c, by Thermo Fisher Scientific (2 mentions)
Cyclic amp elisa kit, by Cayman Chemical (2 mentions)
Paraformaldehyde (pfa), by Merck Group (1 mentions)
Neutral buffered formalin, by Thermo Fisher Scientific (1 mentions)
Proteases inhibitors cocktail, by Merck Group (1 mentions)
Protease inhibitor cocktail, by Cell Signaling Technology (1 mentions)
Micro bicinchoninic acid bca assay, by Thermo Fisher Scientific (1 mentions)
58 protocols using tissue homogenizer
1
Intranasal Infection Assay for Mycobacterium smegmatis
2
Immunohistochemistry and Protein Analysis
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For immunohistochemistry, mice (n = 4−6 per group) were anesthetised with isofluorane and transcardially perfused with ice-cold saline followed by ice-cold 4% paraformaldehyde (Sigma-Aldrich, St Louis, MO, USA) in 0.1 M phosphate-buffered saline. Brains were post-fixed in 10% neutral-buffered formalin (Thermo Fisher Scientific, Mississauga, ON, Canada) for 16 h and transferred to 70% ethanol. Brains were paraffin embedded and sectioned at 4 μm.
For western blot and ELISA (n = 4−8 per group), anesthetised mice were cervically dislocated, the hippocampus and cortex dissected out and immediately frozen on dry ice. Proteins were extracted in 5× volume/weight with radioimmunoprecipitation assay (RIPA) buffer (50 mM Tris-HCl, pH 7.4, 1% NP-40, 150 mM NaCl, 0.25% Na-deoxycholate, 1 mM EDTA, 1 mM Na3VO4, 1 mM NaF, 1 mM PMSF, 10 µl ml−1 of Proteases Inhibitors Cocktail (Sigma-Aldrich)) on ice with a tissue homogenizer (OMNI International, Kennesaw, GA, USA). Samples were centrifuged (20,000 × g, 4 °C) for 20 min, the supernatant recovered, and protein quantified using the BCA method. The RIPA-insoluble pellet was further extracted in 70% formic acid in dH2O, evaporated, and resolubilized in 200 mM Tris-HCl, pH 7.5.
For western blot and ELISA (n = 4−8 per group), anesthetised mice were cervically dislocated, the hippocampus and cortex dissected out and immediately frozen on dry ice. Proteins were extracted in 5× volume/weight with radioimmunoprecipitation assay (RIPA) buffer (50 mM Tris-HCl, pH 7.4, 1% NP-40, 150 mM NaCl, 0.25% Na-deoxycholate, 1 mM EDTA, 1 mM Na3VO4, 1 mM NaF, 1 mM PMSF, 10 µl ml−1 of Proteases Inhibitors Cocktail (Sigma-Aldrich)) on ice with a tissue homogenizer (OMNI International, Kennesaw, GA, USA). Samples were centrifuged (20,000 × g, 4 °C) for 20 min, the supernatant recovered, and protein quantified using the BCA method. The RIPA-insoluble pellet was further extracted in 70% formic acid in dH2O, evaporated, and resolubilized in 200 mM Tris-HCl, pH 7.5.
3
Measuring Testicular TBARS Post-Irradiation
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Thiobarbituric acid reactive substances ( TBARS) productions in testicular germ cells were measured at 2 h, 4h, and 8h post-irradiation. Testes were homogenized in pre-chilled PBS (10 % w/v) using tissue homogenizer (OMNI TH, USA). TBARS level in testicular germ cells were measured following the standard protocol described elsewhere [39 (link)]. TBARS were represented as nmol per mg of protein.
4
Serum and Hepatic Triglyceride Quantification
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The serum TG level was determined using a commercial enzymatic assay kit (Asan Pharmaceutical Co., Seoul, Republic of Korea) according to the manufacturer’s instructions.
The stored liver tissues (100 mg) were homogenized in 1 ml of distilled water using a tissue homogenizer (OMNI International, Warrenton, VA, United States). The homogenate was extracted with 5 ml of a chloroform-methanol (2:1 v/v) mixture and centrifuged at 7,000 g for 5 min. The chloroform layer was aspirated, dried, and resolved by isopropanol. The hepatic TG content was determined using a kit, as mentioned above for serum TG analysis.
The stored liver tissues (100 mg) were homogenized in 1 ml of distilled water using a tissue homogenizer (OMNI International, Warrenton, VA, United States). The homogenate was extracted with 5 ml of a chloroform-methanol (2:1 v/v) mixture and centrifuged at 7,000 g for 5 min. The chloroform layer was aspirated, dried, and resolved by isopropanol. The hepatic TG content was determined using a kit, as mentioned above for serum TG analysis.
5
Extraction of Antigenic Proteins from Strongyloides venezuelensis
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For antigenic extraction, 300.000 S. venezuelensis filariform larvae were resuspended in phosphate-buffered saline (PBS, 0.01 mol/L, pH 7.2) and disrupted in an ice bath using a tissue homogenizer (OMNI International, Kennesaw, USA) with 5 cycles of 5 min each, and then submitted to 8 ultrasound cycles for 20 s at 40 kHz (Thorton, Inspec Eletrônica, São Paulo, Brazil). After an overnight incubation period at 4°C under constant gentle shaking, the suspension was centrifuged at 12 400 g for 30 min at 4°C and the supernatant (saline extract) was quantified [40] , and aliquots were subdivided and stored at −20°C until use on competitive Phage-ELISA.
6
Determination of Cellular dNTP Pools
7
Protein Extraction from Frozen Tissues
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After the mice were euthanized, joint tissues were collected from mice in the CIA model, and lung tissues were obtained from mice in the HDM-challenge model, as described above. The tissues were flash frozen in liquid nitrogen and stored in −80°C until further use. The flash frozen tissues were homogenized on ice using a tissue homogenizer (Omni International, USA), in protein extraction buffer T-PER (Thermo Scientific, USA) containing protease inhibitor cocktail (Cell Signaling Technology, Denver, USA). The homogenates were centrifuged at 10,000 × g, at 4°C for 10 min. The supernatants were collected, aliquoted and stored in −20°C until use. Total protein amount was estimated in the supernatants using micro-Bicinchoninic acid (BCA) assay (Thermo Scientific, USA) according to the manufacturer's instructions.
8
Standardized Intestinal Trehalase Activity
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We sampled intestinal tissues of three bat species, including two Old World fruit bats (C. sphinx and R. leschenaultii) and one insectivorous species (H. armiger). Following the methods of a previous study (Martínez del Rio et al. 1995 ), thawed tissues were homogenized in precooling phosphate buffered saline buffer using an OMNI tissue homogenizer. The substrate trehalose solution (56.0 mM) was prepared using 0.1 M maleate/NaOH buffer (pH = 6.5). Next, 100 µl tissue homogenates were incubated at 37 °C with 100 µl trehalose solutions. After 10 min incubation, 3 ml stop/develop reagents were added to arrest the reactions. The absorbance at 405 nm was evaluated using a spectrophotometer and standardized intestinal trehalase activity was measured as µmol/min/g (Martínez del Rio et al. 1995 ).
9
Adipose Tissue RNA Extraction
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Total RNA was extracted from stromal vascular fraction cells using QIAzol® and isolated via an miRNeasy Mini Kit (Qiagen). For whole adipose tissue, frozen gWAT was disrupted using a tissue homogenizer (Omni International) in the presence of QIAzol®, and total RNA was isolated via an RNeasy Lipid Tissue Mini Kit (Qiagen). Kits were utilized as per the manufacturer's instructions. RNA was quantitated via Qubit, and the quality was assessed via Agilent TapeStation 4200 for RNA‐seq and NanoString applications. The RNA integrity number (RIN) was greater than 7 for all samples analyzed.
10
Quantifying Fibrotic Protein Levels
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