Raw264.7 macrophages and BMDMs (1 × 106 cells/mL) were treated with interferon-γ (IFN-γ) at 20 ng/mL for 2 h. Da-g-Xan at 1, 5, or 20 μg/mL was added, and cells without Da-g-Xan were considered as controls. Twenty-four hours later, cells were collected and incubated with CD11b (0.25 μg/test, Invitrogen, 56-0112-82), CD86 (0.125 μg/test, Invitrogen, 12-0862-82) and CD206 (1 μg/mL, abcam, ab195191). Then, cells were measured using a BD FACSCalibur CellSorting System (BD Bioscience, USA) and analyzed with FlowJo software (Tree Star Inc, USA). This experiment was performed in triplicate. Dissected tissues from rat colonic anastomosis were washed with Hanks Balanced Salt Solution (HBSS) for three times and minced into 1-2 mm pieces. The tissue pieces were digested with 2 mL GEXSCOPETM Tissue Dissociation Solution (Singleron, China), and the red blood cells were removed with 2 mL GEXSCOPETM red blood cell lysis buffer (Singleron, China). The resulting cells were stained with CD 206 (1 μg/mL, abcam, ab125028) with secondary donkey anti-rabbit antibodies (1 μg/mL, abcam, ab150075), CD11b (2 μg/mL, BD Biosciences, cat. 554862), CD86 (2.5 μg/mL, BD Biosciences, cat. 561961) and then measured using the CellSorting System.
Gexscope tissue dissociation solution
Manufactured by Singleron Biotechnologies
Sourced in China, United States
GEXSCOPE Tissue Dissociation Solution is a specialized reagent designed to facilitate the dissociation of tissue samples into single-cell suspensions. The solution contains a proprietary blend of enzymes and buffers optimized for gentle and efficient tissue disaggregation, enabling the release of individual cells from the extracellular matrix and tissue structures.
Automatically generated - may contain errors
Lab products found in correlation
Gexscope red blood cell lysis buffer, by Singleron Biotechnologies (24 mentions)
Gexscope tissue preservation solution, by Singleron Biotechnologies (21 mentions)
Trypan blue, by Merck Group (10 mentions)
Tc20 automated cell counter, by Bio-Rad (9 mentions)
40 micron sterile strainer, by Corning (3 mentions)
40 μm sterile cell strainer, by Corning (2 mentions)
Ab125028, by Abcam (1 mentions)
Ab195191, by Abcam (1 mentions)
Facscalibur cell sorting system, by BD (1 mentions)
Hiseq x10, by Illumina (1 mentions)
Hank s balanced salt solution hbss, by Merck Group (1 mentions)
Macs tissue storage solution, by Miltenyi Biotec (1 mentions)
Hiseq x platform, by Illumina (1 mentions)
Phase contrast light microscope, by Nikon (1 mentions)
Trypan blue solution, by Merck Group (1 mentions)
Sterile cell strainer, by BD (1 mentions)
Python automated tissue dissociation system, by Singleron Biotechnologies (1 mentions)
7 aad, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
39 protocols using gexscope tissue dissociation solution
1
Effects of Da-g-Xan on Macrophage Polarization
2
Bladder Tissue Dissociation and Single-Cell Preparation
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The bladder samples employed in this study were procured from patients who underwent transurethral resection of bladder tumors at Shenzhen Luohu District People's Hospital, in accordance with the guidelines established by the Ethics Committee Board of the aforementioned hospital (2020‐LHORMYY‐KYLL‐007). Written informed consent was obtained from all participants. After surgery, fresh tissue samples were collected and preserved in the GEXSCOPETM Tissue Preservation Solution (Singleron, Cologne, Germany) for subsequent processing. Tissue samples were initially washed with phosphate‐buffered saline (PBS; Gibco, USA), then cut into small pieces of 1–2 mm, and digested using GEXSCOPETM Tissue Dissociation Solution (Singleron) at 37°C with oscillation for 15 min. Following digestion, the samples were passed through a 40‐µm sterile strainer, and the supernatant was removed by centrifugation at 1000 rpm for 5 min at 4°C. The cell pellet was then resuspended in 1 mL PBS, treated with 2 mL GEXSCOPETM red blood cell lysis buffer (Singleron), and incubated on ice for 10 min to lyse the red blood cells. Lastly, the single‐cell suspension was collected, resuspended in PBS, and subjected to further analysis.
3
Single-cell RNA-seq from Fresh Tissues
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Fresh tissue samples were immediately stored upon collection in the GEXSCOPETM Tissue Preservation Solution (Singleron Biotechnol-ogies) within 2–8°C. Before digested in GEXSCOPETM Tissue Dissociation Solution (Singleron Biotechnologies) at 37°C, the specimens were firstly washed with HBSS for three times and grinded into 1–2 mm pieces. After digestion, cell debris was filtered out and the cells were centrifuged at 1000 rpm, 4°C, for 5 minutes and cell pellets were resuspended into 1 ml PBS. To remove red blood cells, 2 mL GEXSCOPETM Red Blood Cell Lysis Buffer (Singleron Biotechnologies) was added to the cell suspension and incubated at 25°C for 10 minutes. The mixture was centrifuged at 1000 rpm for 5 min and the cell pellet was then resuspended in PBS. Single cell suspension was loaded onto a microfluidic chip and single cell RNA-seq libraries were constructed according to the manufacturer’s instructions (Singleron Biotechno-logies). The resulting single cell RNA-seq libraries were sequenced on an Illumina HiSeq X10 instrument with 150bp paired-end reads.
4
Extracting Rat DRG Single Cells
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In total, 24 rats (eight in each group) were sacrificed 28 days after STZ injection. Among these animals, two control group rats, four diabetic rats with MA, and two diabetic rats without MA were used for scRNA-seq. DRGs, dissected from rats of the abovementioned groups, were washed with Hanks Balanced Salt Solution (HBSS; Sigma-Aldrich, USA) three times. Then, the tissue pieces were transferred into a 15 ml centrifuge tube and digested with 2 ml GEXSCOPETM Tissue Dissociation Solution (Singleron, China) at 37°C for 15 min under sustained agitation. After digestion, the samples were filtered using 40 μm sterile strainers and centrifuged at 1,000 rpm for 5 min. The supernatant was discarded and the sediment was resuspended in 1 ml phosphate-buffered saline (PBS; HyClone, USA). Two milliliters of GEXSCOPETM red blood cell lysis buffer (Singleron, China) was added to remove the red blood cells for 10 min at 25°C. The solution was then centrifuged at 500 × g for 5 min and suspended in PBS. The sample was stained with trypan blue (Sigma-Aldrich, USA) and microscopically evaluated.
5
Testis Tissue Dissociation Protocol
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The testes were washed with Hanks Balanced Salt Solution (HBSS) three times and digested in 2 mL of GEXSCOPETM Tissue Dissociation Solution (Singleron Biotechnologies) using a Singleron PythoN™ Automated Tissue Dissociation System (Singleron Biotechnologies) at 28 °C for 15 min. The mixture was then centrifuged at 500 g for 5 min and resuspended with PBS. Finally, the samples were stained with trypan blue (Sigma-Aldrich) and cellular viability was evaluated microscopically.
6
Efficient Tissue Dissociation and Cell Isolation
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Prior to tissue dissociation, the specimens were washed with Hanks balanced salt solution (HBSS) three times and minced into 1–2 mm pieces. The tissue pieces were digested in 2 mL GEXSCOPETM Tissue Dissociation Solution (Singleron Biotechnologies) at 37 °C for 15 min in a 15 mL centrifuge tube with continuous agitation. Following digestion, a 40-micron sterile strainer (Corning) was used to separate cells from cell debris and other impurities. The cells were centrifuged at 1000 rpm and 4 °C for 5 min, and the cell pellet was resuspended in 1 mL PBS (HyClone). To remove red blood cells, 2 mL GEXSCOPETM Red Blood Cell Lysis Buffer (Singleron Biotechnologies) was added to the cell suspension and incubated at 25 °C for 10 min. The mixture was then centrifuged at 1000 rpm for 5 min, and the cell pellet was resuspended in PBS. Cells were counted with a TC20 automated cell counter (Bio-Rad).
7
Single-Cell Sequencing of Tumor Tissue
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The fresh tumor tissue was placed in the cold MACS Tissue Storage Solution (Miltenyi Biotec, Cat: 130-100-008) and was transported to the laboratory at low temperature. GEXSCOPETM Tissue Dissociation Solution (Singleron, China) was used to lysate tissues and digest the cells for single cell suspension, while the Illumina HiSeq X platform (Illumina, San Diego, CA, USA) was used for sequencing.
8
Single-Cell Isolation from Fresh Tissue
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After the fresh tissue samples were surgically removed, they were stored in GEXSCOPETM Tissue Preservation Solution (Singleron, Cologne, Germany) and taken back to the laboratory for further processing. First, the tissue samples were washed by using Hanks Balanced Salt Solution (HBSS), and then cut them into 1~2 mm pieces. The small tissue pieces were transferred to a 15-ml centrifuge tube and digested using GEXSCOPETM Tissue Dissociation Solution (Singleron) under shaking conditions at 37°C for 15 min. After the digestion, the samples were filtered with 40 µm sterile strainers, and then centrifuged at 1,000 rpm at 4°C for 5 min. The supernatant was removed, and the cells were resuspended with 1 ml PBS, then added 2 ml GEXSCOPETM red blood cell lysis buffer (Singleron) and incubated for 10 min to lyse the red blood cells. Finally, the single cell suspension was collected after resuspension with PBS.
9
Isolation of Viable Tumor Cells from Lesions
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Fresh tumor lesions were stored in GEXSCOPETM tissue preservation solution (Singleron Bio Com, Nanjing, China) and processed on ice after the surgery within 30 mins. The specimens were washed with Hanks Balanced Salt Solution (HBSS) three times and minced into 1–2 mm pieces. Then, the tissue pieces were digested with 2 mL of GEXSCOPETM tissue dissociation solution (Singleron) at 37 °C for 15 min with sustained agitation. After digestion, the samples were filtered through 40-µm sterile strainers and centrifuged at 800 × g for 5 min. Subsequently, the supernatants were discarded, and the cell pellets were suspended in 1 mL phosphate-buffered saline (PBS; HyClone, United States). To remove red blood cells, 2 mL of GEXSCOPETM red blood cell lysis buffer (Singleron) was added, and cells were incubated at 25 °C for 10 min. The solution was then centrifuged at 500 × g for 5 min and resuspended in PBS. The samples were stained with trypan blue (Sigma, United States) and the cellular viability was evaluated under the phase contrast light microscope (Nikon, Japan).
10
Single-cell isolation from fresh tumors
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Fresh tumor lesions were stored in cold GEXSCOPETM tissue preservation solution (Singleron Bio Com, Nanjing, China) and transferred for processing on ice within 30 min after resection. The tissue samples were trimmed, washed with Hanks balanced Salt Solution (HBSS) three times and cut into 1–2 mm pieces. Samples were digested using 2 mL of GEXscopetM tissue dissociation solution (Singleron). After digestion, the cell suspension was filtered using a 40-µm sterile strainers and centrifuged at 1000 rpm for 5 min. Dissociated cells were pelleted and resuspended in 1 mL phosphate-buffered saline (PBS; HyClone, United States). Subsequently, red blood Cells were removed with 2 mL of GEXSCOPETM red blood cell lysis buffer (Singleron). The solution was then centrifuged at 500 rpm for 5 min and resuspended in PBS. The samples were dyed with trypan blue solution (Sigma, United States) and observed under the phase contrast light microscope.
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