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Nylon filter

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Nylon filters are a type of laboratory equipment used for filtration purposes. They are made of nylon material and are designed to separate solid particles from liquids or gases, allowing the passage of the desired fluid while retaining the unwanted components.

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Lab products found in correlation

Dnase 1, by Roche (5 mentions) Facsaria 2, by BD (3 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions) Dispase 2, by Roche (2 mentions) Collagenase a, by Roche (2 mentions) Glass backed silica plates, by Merck Group (2 mentions) Ycfac liquid medium, by Anaerobe Systems (2 mentions) Liberase tl, by Roche (2 mentions) Anti f4 80 bm8, by Thermo Fisher Scientific (2 mentions) 7 aminoactinomycin d via probe, by BD (2 mentions) Collagenase d, by Roche (2 mentions) Fc block, by BD (2 mentions) Dnase 1, by Worthington (2 mentions) Anti cd45.2 fluorescein isothiocyanate, by BD (2 mentions) Collagenase type 4, by Merck Group (2 mentions) Rpmi 1640, by Thermo Fisher Scientific (2 mentions) Gentlemacs dissociator, by Miltenyi Biotec (2 mentions) Cellquest software, by BD (2 mentions) Facscalibur, by BD (2 mentions) Streptomycin, by Thermo Fisher Scientific (2 mentions)

25 protocols using nylon filter

1

Isolation and Purification of Murine Male Germ Cells

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All animal studies were approved by the Institutional Animal Care and Use Committee (No: A10871, date of approval: 04/10/2020; No: A10979, date of approval: 03/18/2021) of Fu Jen Catholic University. Murine male germ cells were isolated using a centrifugal system according to the density of different types of germ cells, as described previously.
28 (link) Briefly, testes were decapsulated and the seminiferous tubules were enzymatically digested, after which the germ cell suspensions were filtered through 35‐μM nylon filters (Falcon). The suspension of single cells was centrifuged at different gravity levels on a Kubota centrifuge 3330. Germ cells were collected at different developmental stages. Mature spermatozoa were collected from the cauda epididymides of adult male mice. Finally, suspended male germ cells were spread on a slide and air‐dried for further analysis.
Lin Y., Wang Y., Lai T., Teng J., Lin C., Ke C., Yu I., Lee H., Chan C., Tung C., Conrad D.F., O'Bryan M.K, & Lin Y. (2023). Deleterious genetic changes in AGTPBP1 result in teratozoospermia with sperm head and flagella defects. Journal of Cellular and Molecular Medicine, 28(2), e18031.
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2

Isolation and Cultivation of Lung Fibroblasts

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Mouse lung fibroblasts, MLg (MLg 2908) were purchased from ATCC (Manassas, VA, USA) (CCL-206) and cultivated in DMEM/HAM’s F12 (catalog# E15-813, PAA (Pasching, Austria)) medium containing 10% FBS (PAA). For isolation of primary human lung fibroblasts, specimens from lung lobes or segmental lung resections were dissected into pieces of 1–2 mm2 in size and digested by 5 mg of Collagenase I (Biochrom (Berlin, Germany)) at 37 °C for 2 hours. Subsequently, samples were filtered through nylon filters with a pore size of 70 μm (BD Falcon, (Bedford, MA, USA)). Filtrates, containing the cells, were centrifuged at 400 g, 4 °C for 5 minutes. Pellets were resuspended in DMEM/F-12 medium (Gibco, (Darmstadt, Germany)) supplemented with 20% fetal bovine serum (PAA) and plated on 10 cm cell-culture dishes and subsequently cultured to a confluence of 80–90% in DMEM/HAM’s F12 medium containing 20% FBS. All cells were cultivated under standard conditions (5% CO2 and 37 °C). MLg fibroblasts and primary cells were not used at passage numbers higher than 15 and 10, respectively.
Oehrle B., Burgstaller G., Irmler M., Dehmel S., Grün J., Hwang T., Krauss-Etschmann S., Beckers J., Meiners S, & Eickelberg O. (2015). Validated prediction of pro-invasive growth factors using a transcriptome-wide invasion signature derived from a complex 3D invasion assay. Scientific Reports, 5, 12673.
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3

Enzymatic Dissociation of Murine Hindlimb Muscles

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Hindlimb muscles deriving from a single mouse were dissected and finely minced in Hank’s balanced salt solution (HBSS) (Gibco). Minced muscles were enzymatically digested for 60 min (postnatal muscles) to 90 min (P56 adult muscles) at 37°C with agitation every 20 min in a solution containing collagenase A (2 mg/mL, Roche), dispase II (3 mg/mL, Roche), DNase I (10 μg/mL, Roche), 0.4 mM CaCl2 (Sigma), and 5 mM MgCl2 (Sigma). The resulting cell suspension was washed with HBSS with addition of 0.2% bovine serum albumin (BSA) and 0.5% antibiotics (Gibco) and passed successively through 100- and 40-μm nylon filters (Falcon). Red blood cells were lysed in 1 mL of red blood cell lysing buffer (BD Biosciences) for 20 min. Cells were then transferred to flow cytometry collection tube.
Gattazzo F., Laurent B., Relaix F., Rouard H, & Didier N. (2020). Distinct Phases of Postnatal Skeletal Muscle Growth Govern the Progressive Establishment of Muscle Stem Cell Quiescence. Stem Cell Reports, 15(3), 597-611.
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4

Extraction and Analysis of Secondary Metabolites

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Strains were grown for 24 or 72 hours in YCFAC liquid medium (Anaerobe Systems) containing 100 μM DCA. 2g NaCl2 was added to 7 mL culture supernatant, followed by 700 μL 6M HCl. The supernatant was extracted with 5 mL ethyl acetate followed by dehydration in 2 g MgSO4, and a second extraction with ethyl acetate. The organic extract was filtered through 40 μm nylon filters (Falcon), then dried by vacuum centrifugation in glass tubes. The pellet was resuspended in methanol, tubes were washed in an equal volume of acetone and the solvent was again removed by vacuum centrifugation. The final dried pellet was resuspended in 100 μL acetone and was spotted onto glass-backed silica plates (Sigma), which were developed in 70:20:2 benzene/1,4-dioxane/acetic acid. Plates were stained in 10% w/v CuSO4 8% H3PO4 and dried over a hot plate.
Campbell C., McKenney P.T., Konstantinovsky D., Isaeva O.I., Schizas M., Verter J., Mai C., Jin W.B., Guo C.J., Violante S., Ramos R.J., Cross J.R., Kadaveru K., Hambor J, & Rudensky A.Y. (2020). Bacterial metabolism of bile acids promotes peripheral Treg cell generation. Nature, 581(7809), 475-479.
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5

Extraction and Analysis of Secondary Metabolites

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Strains were grown for 24 or 72 hours in YCFAC liquid medium (Anaerobe Systems) containing 100 μM DCA. 2g NaCl2 was added to 7 mL culture supernatant, followed by 700 μL 6M HCl. The supernatant was extracted with 5 mL ethyl acetate followed by dehydration in 2 g MgSO4, and a second extraction with ethyl acetate. The organic extract was filtered through 40 μm nylon filters (Falcon), then dried by vacuum centrifugation in glass tubes. The pellet was resuspended in methanol, tubes were washed in an equal volume of acetone and the solvent was again removed by vacuum centrifugation. The final dried pellet was resuspended in 100 μL acetone and was spotted onto glass-backed silica plates (Sigma), which were developed in 70:20:2 benzene/1,4-dioxane/acetic acid. Plates were stained in 10% w/v CuSO4 8% H3PO4 and dried over a hot plate.
Campbell C., McKenney P.T., Konstantinovsky D., Isaeva O.I., Schizas M., Verter J., Mai C., Jin W.B., Guo C.J., Violante S., Ramos R.J., Cross J.R., Kadaveru K., Hambor J, & Rudensky A.Y. (2020). Bacterial metabolism of bile acids promotes peripheral Treg cell generation. Nature, 581(7809), 475-479.
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6

Isolation and Sorting of Murine Lung Cells

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Mouse lungs were digested with 10 mg of collagenase (Fujifilm Wako), 10 mg of dispase (Gibco), and 10 μg of DNase (Worthington Biochemical, Lakewood, NJ) in 10 mL of serum-free DMEM at 37 °C for 45 min. The cells were resuspended in PBS-1% BSA and sequentially filtered through 100- and 40-μm nylon filters (Falcon). After treatment with red blood cell lysis buffer (Sigma-Aldrich), cells were resuspended in PBS-1% BSA and loaded onto a cell sorter (FACSAria III, BD Biosciences). For FACS sorting, cells were labeled with the following antibodies: anti-CD144-PE/Cy7 (1:200, 138015, BioLegend), anti-CD11b-APC (1:200, 101211, BioLegend), and anti-F4/80-FITC (1:400, 123107, BioLegend). FACS data were analyzed using FACS Diva 8.0.2 (BD) and Flowjo v7.6 (Flowjo LLC).
Han Y., Tomita T., Kato M., Ashihara N., Higuchi Y., Matoba H., Wang W., Hayashi H., Itoh Y., Takahashi S., Kurita H., Nakayama J., Okumura N, & Hiratsuka S. (2023). Citrullinated fibrinogen-SAAs complex causes vascular metastagenesis. Nature Communications, 14, 4960.
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7

Isolation of Murine Satellite Cells

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Satellite cells were isolated from skeletal muscles of 4-week-old mice by FACS, after enzymatic dissociation with a solution of CMF containing 0.02% BSA (Bovine Serum Albumin) (Sigma-Aldrich), 1% of Penicillin/Streptomycin (Sigma-Aldrich), 0.25 mg/ml Dispase II (Roche); 2 μg/μl Collagenase A (Roche), 8 mM CaCl2,10 ng/μl DNase I (Roche) and 5 mM MgCl2 for 2 hours at 37 °C. Cell suspensions were filtered through a 100-μm and a 40-μm nylon filters (Falcon) and stained with primary antibodies for 30 min on ice. The following antibodies were used: CD45- eFluor450 (eBioscience), Ter119- eFluor450 (eBioscience), CD31- Pacific Blue (Invitrogen) and Sca1-FITC (BD Bioscience), integrin- α7–APC (Ab-Lab). Flow cytometry analysis and cell sorting were performed on a DAKO-Cytomation MoFlo High Speed Sorter.
Marroncelli N., Bianchi M., Bertin M., Consalvi S., Saccone V., De Bardi M., Puri P.L., Palacios D., Adamo S, & Moresi V. (2018). HDAC4 regulates satellite cell proliferation and differentiation by targeting P21 and Sharp1 genes. Scientific Reports, 8, 3448.
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8

Tumor and Lymph Node Dissociation Protocol

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LNs and primary tumors were mechanically dissociated with forceps and scalpels and placed in DMEM containing 2 mg/ml collagenase (Sigma) for 20 min at 37°C (for LN metastatic burden evaluation, collagenase incubation time was increased to 40 min). The suspensions were incubated in 0.1 mg/ml DNase I (Roche) for 10 mins. Tissue suspensions were resuspended with P1000, filtered through 40 μm nylon filters (Falcon), and aliquoted for total cell counting to adjust equal cell numbers for antibody staining. For antibody staining and FACS analysis, cells were fixed in 1 × Cytofix fixation buffer (BD) for 10 min at 37°C. After washing with PBS, cells were permeabilized in 0.1% Triton X-100 in PBS for 5 min at room temperature. Cells were washed and resuspended in staining buffer (BD) and then blocked with mouse FcR blocking reagent. Then samples were incubated with FACS antibodies and processed as described in the previous section.
Singh R, & Choi B.K. (2019). Siglec1-expressing subcapsular sinus macrophages provide soil for melanoma lymph node metastasis. eLife, 8, e48916.
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9

Peritoneal Exudate Cells Isolation

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Axenic M. corti tetrathyridia were washed 3 times and 60 larvae in 1 ml PBS were injected i.p. in Balb/c mice. Control mice received 60 dead larvae (heat killed by treatment at 100°C for 15 min) or 1 ml of PBS as a mock control. At day 3 and 7 p.i., mice were sacrificed by CO2 asphyxiation and peritoneal exudate cells were collected by flushing the peritonea with 5 ml of complete medium i.e. RPMI 1640 (Biochrom, Berlin, Germany) supplemented with 10% heat-inactivated fetal calf serum (Biochrom, Berlin, Germany), 100 U/mL of penicillin (Biochrom, Berlin, Germany), 100 μg/mL of streptomycin (Biochrom, Berlin, Germany), 2 mM L-glutamine (Sigma, St. Louis, USA) and 50 μM 2-mercaptoethanol (Merck, Darmstadt, Germany). The suspensions of peritoneal cells were sieved through a 40 μM nylon filters (BD Biosciences, Durham, NC, USA). Red blood cells were lysed using 1.4% NH4Cl for 5 min at 37°C and then washed with the complete RPMI medium. Viable cells were counted using a Neubauer chamber by trypan blue exclusion.
Vendelova E., Camargo de Lima J., Lorenzatto K.R., Monteiro K.M., Mueller T., Veepaschit J., Grimm C., Brehm K., Hrčková G., Lutz M.B., Ferreira H.B, & Nono J.K. (2016). Proteomic Analysis of Excretory-Secretory Products of Mesocestoides corti Metacestodes Reveals Potential Suppressors of Dendritic Cell Functions. PLoS Neglected Tropical Diseases, 10(10), e0005061.
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10

Isolation and Characterization of T Cell Subsets

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T cells were magnetically isolated from pooled lymph node and spleen cells using CD4+ negative isolation kits from either EasySep or Miltenyi according to the manufacturer’s instructions. Naïve (CD25−CD62LhiCD44lo) or memory (CD25−CD62LloCD44hi) T cells were further sorted for CCR6 expression by FACS sorting (FACS Aria II; BD). Cells were ≥ 98% pure following isolation as determined by FACS analyses. CNS-infiltrating mononuclear cells were isolated from brain and spinal cord after intracardiac perfusion with PBS. Tissue was minced and digested with 0.33mg/ml Liberase TL and 20U/ml DNase (Roche) for 30 min at 37 °C. Digested tissue was forced through 70μm nylon filters (BD). Mononuclear cells were isolated from the interface following 30%/70% Percoll (Sigma) gradient centrifugation.
Carlson T.J., Pellerin A., Djuretic I.M., Trivigno C., Koralov S.B., Rao A, & Sundrud M.S. (2014). Halofuginone-induced amino acid starvation regulates Stat3-dependent Th17 effector function and reduces established autoimmune inflammation. Journal of immunology (Baltimore, Md. : 1950), 192(5), 2167-2176.
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