PBMCs were separated by standard Ficoll-Paque (GE Healthcare, Uppsala, Sweden) density gradient centrifugation, and then cryopreserved in FBS with 10% DMSO (Sigma-Aldrich, St. Louis, MO, USA) until use. To purify single-cell suspensions of lymphocytes from tissue samples, aortic tissue was placed in a gentle MACS C-Tube (Miltenyi Biotec, Bergisch Gladbach, Germany) enzyme H, enzyme R, and enzyme A (Miltenyi Biotec) pre-mixed in RPMI. These C-tubes were then placed on a gentle MACS Octo Dissociator. Next, the cell suspension was passed through a 70-μm cell strainer, and the cells were washed once and then resuspended in media. The number and viability of mononuclear cells assessed using a Cellometer® Auto 2000 (Nexcelom, Lawrence, MA, USA). The cells were then washed once and cryopreserved. CD45+ cells from aortic tissue were purified from single-cell isolates using the REAlease CD45 (TIL) MicroBead Kit (130-121-563; Miltenyi Biotec).
Macs c tube
Manufactured by Miltenyi Biotec
Sourced in Germany, United States, Italy
The MACS C tube is a laboratory equipment designed for cell separation and isolation. It is a component of Miltenyi Biotec's magnetic-activated cell sorting (MACS) technology. The MACS C tube facilitates the separation of target cells from a heterogeneous cell population using magnetic beads conjugated with specific antibodies.
Automatically generated - may contain errors
Lab products found in correlation
Macs dissociator, by Miltenyi Biotec (8 mentions)
Gentlemacs dissociator, by Miltenyi Biotec (5 mentions)
Tumor dissociation kit, by Miltenyi Biotec (5 mentions)
L1825 bc, by Merck Group (3 mentions)
Dead cell removal kit, by Miltenyi Biotec (3 mentions)
Lysis buffer, by Thermo Fisher Scientific (3 mentions)
Ficoll paque, by GE Healthcare (2 mentions)
Dmem and ham s f12 media mixture, by Thermo Fisher Scientific (2 mentions)
Macs octo dissociator with heaters, by Miltenyi Biotec (2 mentions)
Ficoll, by Solarbio (2 mentions)
Ficoll, by TBD Science (2 mentions)
Rpmi 1640, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Realease cd45 til microbead kit, by Miltenyi Biotec (1 mentions)
Cellometer auto 2000, by Revvity (1 mentions)
Dnase 1, by Merck Group (1 mentions)
Hyaluronidase, by Merck Group (1 mentions)
Liberase, by Merck Group (1 mentions)
Collagenase type 2, by Thermo Fisher Scientific (1 mentions)
32 protocols using macs c tube
1
Isolation and Cryopreservation of PBMCs and CD45+ Cells from Aortic Tissue
2
Tissue Dissociation and Cell Isolation from Ear Samples
3
Tumor Tissue Dissociation and Cell Isolation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tumor tissues were obtained within 1 to 2 h after surgical removal, washed in sterile Dulbecco’s phosphate-buffered saline (PBS) (L1825-BC – Merck Millipore, Italy), and mechanically minced into small pieces (2 to 4 mm). Minced samples were digested using a tumor dissociation kit in a disposable gentle MACS™ C-Tube (Miltenyi Biotec, Italy) according to the manufacturer’s instructions. Samples were digested for 60 min at 37°C in a gentle MACS Octo dissociator, filtered through 70-μm sterile cell strainers, centrifuged at 300 × g for 5 min, and resuspended in a DMEM and HAM’S F12 media mixture (2:1) (Gibco) containing 50 IU/mL penicillin-streptomycin and 4 mM glutamine. Viable cells were counted using an optic phase-contrast microscope [7 (link)].
4
Tumor-Initiating Cell Frequency Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SCID mice bearing Kuramochi xenografts from the above tumour initiation experiment were treated with PBS or ALM201 until tumours reached geometric mean diameter (GMD) of 12 mm3. Tumours were excised, disaggregated using a scalpel and added to a MACs C tube (Miltenyi Biotec, UK) containing collagenase type II (Invitrogen, UK), DNAase type 1 (Sigma-Aldrich, UK) in RPMI/1% penicillin/streptomycin (Invitrogen, UK). Tumours were minced by using a gentleMACS dissociator (Miltenyi Biotec, UK) and incubated at 37 °C in an orbital incubator for 45 min. The cell suspension was resuspended in red blood cell lysis buffer (Roche, UK) for 2–3 min. The cells were resuspended in ice-cold PBS and counted using a haemocytometer. Cells were implanted intradermally, as described above, into secondary SCID mice at 2.5 × 106, 1 × 106, 5 × 105, 1 × 105 and 1 × 104 cells per mouse. Mice did not receive treatment and were observed for tumour initiation for 6 months. The tumour- initiating cell frequency was calculated by using ELDA software.30 (link)
5
Isolation of Peripheral Blood Mononuclear Cells and Tumor-Infiltrating Immune Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We employed Ficoll (Cytiva, USA) density gradient centrifugation to isolate PBMCs from whole blood. Single-cell suspensions were prepared from fresh tumor tissue using a combination of mechanical dissociation, following the manufacturer's guidelines with a gentle MACS C tube (Milteny Biotec, Bergisch Gladbach, Germany), and enzymatic hydrolysis using a tumor dissociation kit (Milteny Biotec). Following the digestion process, the cells were filtered through a 70 µm mesh, subjected to centrifugation with Ficoll (Cytiva, USA), and subsequently, monocytes were resuspended in RPMI-1640.
6
Isolation of Immune Cells from HCC Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Clinical samples were obtained from a prospective cohort of 79 patients with pathologically confirmed HCC who underwent surgical resection at Asan Medical Center (Seoul, Korea) between April 2016 and April 2019. Fresh tumor tissue, adjacent nontumor liver tissue, and whole‐blood samples were collected. Table 1 summarizes the clinical characteristics of the study patients. We also obtained fresh tumor tissues from patients with other cancer types, and the corresponding data are summarized in Supporting Table S1 . All included patients provided written informed consent, and this study was approved by the institutional review board.
Peripheral blood mononuclear cells (PBMCs) were isolated from whole blood by standard Ficoll‐Paque (GE Healthcare, Uppsala, Sweden) density gradient centrifugation. Intrahepatic lymphocytes (IHLs) and TILs were isolated from tissue samples as described.18 Briefly, tissue samples were cut into small pieces (2‐4 mm) and then transferred to a gentle MACS C‐Tube containing a mixture of enzymes (Milteny Biotec, Bergisch Gladbach, Germany). Next, samples were mechanically homogenized and enzymatically digested using the gentle MACS Octo Dissociator (Milteny Biotec). The resulting cell suspension was passed through a 70‐μm cell strainer, and then the cells were washed and cryopreserved.
Peripheral blood mononuclear cells (PBMCs) were isolated from whole blood by standard Ficoll‐Paque (GE Healthcare, Uppsala, Sweden) density gradient centrifugation. Intrahepatic lymphocytes (IHLs) and TILs were isolated from tissue samples as described.
7
Isolation of Tumor Cells from Fresh Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Freshly obtained tumor tissues (within 1–2 h after surgical removal) were washed in sterile Dulbecco's phosphate-buffered saline (PBS) (L1825-BC—Merck Millipore) and mechanically minced into small pieces (2–4 mm). Minced samples were digested using a tumor dissociation kit in a disposable gentle MACS™ C-Tube (Miltenyi) according to the manufacturer's instructions. Samples were digested for 60 min at 37°C in a gentle MACS Octo dissociator and filtered through 70-μm sterile cell strainers, centrifuged at 300 × g for 5 min, and resuspended in a mixture of Dulbecco's modified Eagle medium (DMEM) and Ham's F12 media (2:1) (Gibco) containing 50 IU/ml penicillin–streptomycin and 4 mM glutamine. Finally, viable cells were counted using an optic phase contrast microscope.
8
Isolation of Tumor Cells from Surgical Specimens
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Surgical specimens were retrieved 1 to 2 h after surgery, washed in 50 mL sterile Falcon with Dulbecco’s phosphate-buffered saline (D-PBS) (L1825-BC—Merck Millipore, Italy), and mechanically minced into small pieces (2 mm to 4 mm). Samples were digested for 60 min at 37 °C in a gentle MACS Octo dissociator according to the manufacturer’s instructions, with Milteny tumor dissociation in a MACS™ C-Tube (Miltenyi Biotec, Italy). The cell suspension was then filtered through 70-μm sterile cell strainers, centrifuged at 300 × g for 5 min, and resuspended in a DMEM and HAM’S F12 media mixture (2:1) (Gibco) containing 50 IU/mL penicillin-streptomycin and 4 mM glutamine. Viable cells were counted using an optic phase-contrast microscope.
9
Mouse Brain Dissociation and Cell Sorting
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Whole mouse brains were dissected from adult female CKp25 mice and washed in cold PBS with Ca2+ and Mg2+ (14287080; Thermo Fisher Scientific), placed on a Petri dish, finely diced with a razor, transferred to a MACS C Tube (130-093-237; Miltenyi), treated with the Adult Brain Dissociation Kit, mouse and rat (130-107-677; Miltenyi), and digested with a gentleMACS Octo Dissociator with Heaters (130-096-427; Miltenyi) for 30 min at 37°C. The slurry was then strained through a MACS 70 µm SmartStrainer (130-098-462; Miltenyi). Debris removal solution was then added, followed by a 3,000 ×g, 10 min, 4°C spin, discarding of two upper phases, and PBS wash step 1,000g, 10 min, 4°C spin. Cells were then labeled with CD11b antibody (130-110-554, 1:25 dilution; Miltenyi) and DAPI (D9542-1MG; Millipore) for 40 min followed by sorting into PBS with 1% BSA (700-100P; Gemini). Cells were then pelleted and subject to mRNA extraction using the RNeasy Plus Mini Kit (74134; Qiagen).
10
Kidney Cortex Cell Isolation and Stimulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cortex samples from human kidney were sliced into ∼30-mm3 pieces and digested for 30 min at 37 °C with agitation in a digestion solution containing 25 μg/mL Liberase TM (Roche) and 50 μg/mL DNase (Sigma) in RPMI-1640. Following incubation, samples were transferred to a gentle MACS C Tube (Miltenyi Biotec) and processed using a gentleMACS dissociator (Miltenyi Biotec) on program spleen 4 and subsequently lung 2. The resulting suspension was passed through a 70-μm cell strainer (Falcon) and washed with PBS before leukocyte enrichment using a Percoll density gradient (Sigma). Cells were counted using a hemocytometer with trypan blue. A total of 5 × 105 cells per well were stimulated with Ova or Ova-IC with or without the presence of 2DG (5 mM/mL) for 12 h. Cells were then lysed and processed for RNA extraction and qPCR.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!