Chick embryo livers and lungs were thawed, rinsed with 5 mL PBS and homogenized with a handheld homogenizer (Omni International) using a sterile tip for each sample. DNA extraction was performed using the QIAgen DNeasy Blood & Tissue Kit (QIAGEN Group), following the manufacturer’s specifications. Purified DNA was quantified using a spectrophotometer, adjusted to a concentration of 0.2ug/uL, and stored at -20 degrees C. Quantitative PCR (qPCR) was performed on the DNA samples using primers specific for a human Alu sequence. Alu sequences are primate-specific, and thus their detection in chick organs represents disseminated human cells, or cancer metastases. The copy threshold, or CT, values of the liver and lung specimens from the eight HNSCC cell lines were compared to the CT values of the negative controls of the respective organ. Student T-tests were used to compare the CT values of both organs of each cell line to the respective negative control.
Handheld homogenizer
Manufactured by Omni International
Sourced in United States
The Handheld homogenizer is a laboratory equipment used for the mixing and dispersion of samples. It provides a rapid and efficient method for homogenizing a variety of materials, including liquids, semisolids, and suspensions.
Automatically generated - may contain errors
Lab products found in correlation
Dneasy blood tissue kit, by Qiagen (1 mentions)
Stripping buffer, by Thermo Fisher Scientific (1 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
Nitrocellulose, by Bio-Rad (1 mentions)
Ecl reagent, by GE Healthcare (1 mentions)
Polyclonal antibodies, by Abcam (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
Tnf alpha duoset elisa kit, by R&D Systems (1 mentions)
Ripa buffer, by Thermo Fisher Scientific (1 mentions)
Bicinchoninic acid protein assay, by Thermo Fisher Scientific (1 mentions)
A32957, by Thermo Fisher Scientific (1 mentions)
A32955, by Thermo Fisher Scientific (1 mentions)
Lysis buffer, by Promega (1 mentions)
Rotor gene 3000 instrument, by Qiagen (1 mentions)
Balb c mice, by Charles River Laboratories (1 mentions)
Complete protease inhibitor, by Roche (1 mentions)
11 protocols using handheld homogenizer
1
Detecting Human Cell Metastasis in Chick Organs
2
Western Blot Analysis of Intestinal Tumors
3
Immunological Profiling of Host Tissues
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Specimens from host spleen, lung and colon were mechanically disrupted and homogenized using a handheld homogenizer (Omni International, Kennesaw, GA). Supernatants were collected for ELISA of tumor necrosis alpha (TNF-α), interferon gamma (IFN-γ), RANTES/CCL-5 and phosphorylated STAT 5A/B (eBioscience, San Diego, CA). Inducible nitric oxide synthase (iNOS) levels were detected using polyclonal antibodies (Abcam, Cambridge, UK) for western blotting. Total protein concentration in the supernantant was measured using Bradford’s reagent. Supernatants were then subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Proteins were then transferred from the gel to nitrocellulose membranes and ECL reagent was used for visualization of the protein (GE Healthcare, Buckinghamshire, UK).
4
Rat Brain Tissue Homogenization
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The frontal cortex and hippocampus from each rat were homogenized in ice-cold medium of 1.15% KCl (pH 7.4) to give a 10% (w/v) mixture using a hand-held homogenizer (Omni International, Kennesaw, GA, USA). The supernatants were obtained by centrifugation at 1000× g for 15 min and were used for further analyses.
5
ELISA Quantification of Tumor Necrosis Factor
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The amount of tumor necrosis factor TNFα protein in frozen samples was measured using a TNF-alpha DuoSet ELISA kit (R&D Systems, Minneapolis, Minn.) according to manufacturer protocol. For frozen sample preparation, approximately 500 mg were minced and mixed with 2 mL PBS with protease inhibitor cocktail (Sigma, St. Louis, Mo.). Tissues were homogenized using a hand-held homogenizer (Omni International, Kennesaw, Ga.) on ice. The resulting suspension was centrifuged for 15 min at 1500 × g to collect the lysate.
6
Total Tissue Lysate Preparation
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Total tissue lysates were prepared in RIPA buffer (89901, Thermo Scientific, Waltham, MA) containing protease (A32955, Thermo Scientific, Waltham, MA) and phosphatase (A32957, Thermo Scientific, Waltham, MA) inhibitors. Tissue was homogenized using handheld homogenizer (Omni International TH, Kenasaw, GA) and centrifuged at max speed (12,700 rpm) for 20 min. Aliquots were prepared and stored − 80 °C. The total protein concentration in aliquots of skeletal muscle and liver homogenate was determined using the bicinchoninic acid protein assay (23225, Thermo Fisher Scientific, Waltham, MA) as per manufacturer’s instructions.
7
Hepatic Tissue Homogenate Analysis
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Hepatic tissue homogenates (10% w/v) were prepared in chilled 1.15% KCl at pH 7.4 using a handheld homogenizer (Omni international, Kennesaw, GA, USA). The homogenates were further analyzed for HO-1 (BioVision, Milpitas, CA, USA, # E4525-100) and TIMP-1 (BioVision, Milpitas, CA, USA, # E4844-100) following the manufacturers’ protocols.
8
Tissue Homogenization and Luciferase Assay
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Tissues were removed and placed in flat-bottomed 2.0-mL small capped tubes (USA Scientific, Ocala, FL) with lysis buffer (Promega, Madison, WI) and placed in an ice slush. Tissues were homogenized with a hand-held homogenizer (Omni International, Marietta, GA). Specimens were centrifuged at 4°C at 13,200 rpm for 5 min and then returned to ice. A luminometer (Berthold Detection Systems, Oak Ridge, TN) was used to measure total light emission according to the manufacturer’s instructions. All samples were measured for 10 s after a 5-s delay. Luciferase levels were determined in triplicate by luminometry and recorded as relative light units (RLU).51 (link)
9
Curcumin Extraction and Analysis in Plasma and Tissues
10
Quantification of Kv Channel mRNA Levels
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The samples stored in RNAlater™ were homogenized with a handheld homogenizer (Omni International Inc.). RNA from the tissue homogenates and from cultured VSMCs was isolated with TRIzol Reagent and reverse transcribed as previously described. 16 (link) The mRNA levels of Kv1.3 (Kcna3)), Kv1.5 (Kcna5) and Glyceraldehyde 3-phosphate dehydrogenase (Gapdh) were determined by real-time qPCR using SybrGreen (for Gapdh) or Taqman ® probes (for Kcna3 and Kcna5) on a Rotor-Gene 3000 instrument (Corbett Research) using the 2 -ΔΔCt relative quantification method. 19 (link) mRNA expression levels were normalized to the internal control, Gapdh mRNA. The relative abundance of the genes was calculated from 2 (-ΔCt) , where ΔCt=Ctgenel-CtGapdh and the changes in expression between the different experimental conditions were calculated from 2 (-ΔΔCt) , where ΔΔCt = ΔCt(experimental)-ΔCt(control), the calibrator sample is indicated in each case. In this way, data in the different experimental conditions are presented as the fold change in gene expression. For the representation of these data, the logarithm of 2-ΔΔCt was used, so that a value of 0 means no change, positive values represent increased expression and negative values decreased expression. Total RNA from human brain (BD Biosciences) was used as positive control. Primers sets employed are listed in Supplemental table 1.
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