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Cell tak

Manufactured by Corning
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Sourced in United States, United Kingdom, Germany

Cell-Tak is a specialized adhesive used for attaching cells to various surfaces during cell culture experiments. It is composed of polyphenolic proteins derived from marine mussels. Cell-Tak provides a versatile and non-toxic means of securing cells to a wide range of materials, enabling researchers to maintain cell attachment and viability during experiments.

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Lab products found in correlation

Oligomycin, by Merck Group (29 mentions) Seahorse xfe96 analyzer, by Agilent Technologies (25 mentions) Antimycin a, by Merck Group (22 mentions) Rotenone, by Merck Group (22 mentions) Xfe96 analyzer, by Agilent Technologies (13 mentions) Fluoro carbonyl cyanide phenylhydrazone (fccp), by Merck Group (12 mentions) Oligomycin, by Agilent Technologies (12 mentions) Xfe96 extracellular flux analyzer, by Agilent Technologies (12 mentions) Xf24 extracellular flux analyzer, by Agilent Technologies (11 mentions) Wave software, by Agilent Technologies (10 mentions) Seahorse xf96 analyzer, by Agilent Technologies (10 mentions) Xf96 extracellular flux analyzer, by Agilent Technologies (10 mentions) Seahorse xf cell mito stress test kit, by Agilent Technologies (9 mentions) Seahorse xfe96 extracellular flux analyzer, by Agilent Technologies (9 mentions) Mito stress test kit, by Agilent Technologies (8 mentions) Superfrost plus slide, by Thermo Fisher Scientific (8 mentions) L glutamine, by Merck Group (7 mentions) Seahorse xfp analyzer, by Agilent Technologies (7 mentions) Seahorse xf rpmi medium, by Agilent Technologies (6 mentions) Glutamax, by Thermo Fisher Scientific (6 mentions)

390 protocols using cell tak

1

Coating Glass and Mica with Cell-Tak

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Cell-Tak glass slides were prepared by pipetting 285 ml 100 mM NaHCO3 (pH 8) onto high precision cover glass (24x24 mm2) then 10 μl of Cell-Tak (Corning, 5% (w/v) in acetic acid) and 5 μl of 1 M NaOH. This was covered and left for 20 minutes. The slide was then washed five times with HPLC grade water. The same process with proportionately smaller volumes was used for coating freshly cleaved mica.
Pasquina-Lemonche L., Burns J., Turner R.D., Kumar S., Tank R., Mullin N., Wilson J.S., Chakrabarti B., Bullough P.A., Foster S.J, & Hobbs J.K. (2020). The Architecture of the Gram Positive Bacterial Cell Wall. Nature, 582(7811), 294-297.
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2

Evaluating TLR7/8 Agonists in Murine BMDMs

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BM cells from B6.huTLR8tg mice or C57BL/6 littermates were plated in triplicates in 96-well plates stimulated with TLR7-specific agonist CL264 or TLR8-specific agonist TL-506 (InvivoGen) at the indicated concentrations for 24 h and then TNFα from the supernatants was measured by ELISA (R and D). 2 × 105 BMDM from TLR7-deficient mice were plated overnight in XF media on Seahorse culture plates that were pre-coated with Cell-Tak (Corning cell and tissue adhesive) for 20 min at RT, washed with H2O and air dried before cell plating. Seahorse culture plates were pre-coated with Cell-Tak (25 μl of 11.2 µg/mL) for 20 min at RT (Corning cell and tissue adhesive), washed with H2O, and air dried before cell plating. Cells were stimulated with agonists CL075 (3M002; InvivoGen) or TL-506 (InvivoGen) for 24 h. BMDM extracellular acidification rate was then measured using a Seahorse XF96 Extracellular Flux Analyzer and the Glycolysis Stress Test kit according to manufacturer’s instructions (Agilent). Data were normalized to the cell number.
Maria N.I., Papoin J., Raparia C., Sun Z., Josselsohn R., Lu A., Katerji H., Syeda M.M., Polsky D., Paulson R., Kalfa T., Barnes B.J., Zhang W., Blanc L, & Davidson A. (2023). Human TLR8 induces inflammatory bone marrow erythromyeloblastic islands and anemia in SLE-prone mice. Life Science Alliance, 6(10), e202302241.
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3

Measuring Cellular Respiration Dynamics

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Basal oxygen consumption rate was measured using a Mito Stress test Kit and XF96 Extracellular Flux Analyzer (Seahorse Bioscience), according to manufacturer’s instructions. Cells were resuspended in XF Assay Medium (Seahorse Bioscience) supplemented with 10mM glucose, 1mM pyruvate, and 2mM glutamine. 200,000 cells/well were plated in the XF96 plates pre-coated with Cell-Tak (Corning) and centrifuged to immobilize cells on the bottom of the wells. The plate was incubated in a non-CO2 incubator at 37°C for 2hr to equilibrate. OCR measu rements, taken every 6min, were collected at baseline and after the sequential addition of oligomycin 1uM (final concentration), FCCP 0.5uM, and rotenone 0.75uM + antimycin A 1.5uM.
TMPD-driven oxygen consumption rate was measured using the XF96 Extracellular Flux Analyzer (Seahorse Bioscience). Cells were resuspended in XF Assay Medium (Seahorse Bioscience), not supplemented. 200,000 cells/well were plated in the XF96 plates pre-coated with Cell-Tak (Corning) and centrifuged to immobilize cells on the bottom of the wells. The plate was incubated in a non-CO2 incubator at 37°C for 2hr to equilibrate. OCR measu rements, taken every 6min, were collected at baseline and after the sequential addition of TMPD 0.5mM + ascorbate 10mM + antimycin A 2uM + ADP 1mM, oligomycin 2uM, and azide 20mM.
Lin K.H., Xie A., Rutter J.C., Ahn Y.R., Lloyd-Cowden J.M., Nichols A.G., Soderquist R.S., Koves T.R., Muoio D., MacIver N.J., Lamba J., Pardee T.S., McCall C.M., Rizzieri D.A, & Wood K.C. (2019). Systematic dissection of the metabolic-apoptotic interface in AML reveals heme biosynthesis to be a regulator of drug sensitivity. Cell metabolism, 29(5), 1217-1231.e7.
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4

Metabolic Profiling of Bone Marrow-Derived Cells

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Metabolic effect of stimulation of BMDCs was measured using two assays. For measurement of oxidative phosphorylation, the oxygen consumption rate (OCR) was analyzed by performing an overnight stimulation of BMDCs with either 12.5 μg/mL of PBC micelles, 1 μg/mL of LPS, or untreated control, was carried out in 5 mL polypropylene tubes. 2.5 × 105 stimulated BMDCs were seeded into Cell-Tak (Corning, Corning NY) coated Seahorse plates and a mitochondrial stress test (MST) was conducted according to manufacturer’s specifications using kit concentrations of 1 μM oligomycin, 2 μM FCCP, and 0.5 μM rotenone and antimycin (Agilent, Santa Clara, CA).
For measurement of acute metabolic responses and glycolysis (demonstrated by extracellular acidification rate, ECAR) upon stimulation, untreated BMDCs were seeded into Cell-Tak (Corning, Corning NY) coated seahorse plates at a density of 2.5 × 105 untreated BMDCs per well. Baseline metabolic activity readings were measured, and wells were stimulated with 12.5 μg/mL of micelle, 1 μg/mL of LPS, or medium control, and metabolic measurements are taken over the course of the assay. All metabolic phenotyping was conducted on a Seahorse XFe24 (Agilent, Santa Clara, CA).
Senapati S., Darling R.J., Loh D., Schneider I.C., Wannemuehler M.J., Narasimhan B, & Mallapragada S.K. (2019). Pentablock Copolymer Micelle Nanoadjuvants Enhance Cytosolic Delivery of Antigen and Improve Vaccine Efficacy while Inducing Low Inflammation. ACS biomaterials science & engineering, 5(3), 1332-1342.
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5

Oligodendrocyte Isolation and Culture

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Oligodendrocytes from P6–P8 mice were isolated from wild type as outlined above. The oligodendrocytes were plated on Seahorse XFe96 cell culture microplates (Agilent, Santa Clara, CA) that were pretreated for 20 minutes with Cell-Tak (Corning Inc, Corning, NY) at a concentration of 3.5 μg Cell-Tak/cm2 of surface area. The oligodendrocytes were cultured for 24 hours in complete culture medium containing Dulbecco’s modified Eagle’s medium (DMEM, Gibco, Carlsbad, CA), 10% FBS (Gibco, Carlsbad, CA), 100 U/mL of penicillin, 100 μg/mL of streptomycin, and 0.5 % BSA in PBS, 100 U/mL of IFN-γ (R&D Systems, Minneapolis, MN), or 100 U/mL of LIF R&D Systems, Minneapolis, MN).
Minchenberg S.B, & Massa P.T. (2017). The control of oligodendrocyte bioenergetics by interferon-gamma (IFN-γ) and Src homology region 2 domaincontaining phosphatase-1 (SHP-1). Journal of neuroimmunology, 331, 46-57.
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6

Mitochondrial Stress Test for BMDCs

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For mitochondrial stress tests (MST), BMDCs were stimulated overnight with 0.5 μg/mL CDNs, 1 μg/mL LPS or MPLA, 5 μg/mL imiquimod, CpG ODN, or no stimulation control in 5 mL polypropylene tubes (to avoid cell adherence). Treated BMDCs were seeded into 24 well seahorse plates coated with Cell-Tak (Corning, Corning NY) at a density of 2.5 × 105 cells per well. Mitochondrial stress test was carried out according to manufacturer’s MST protocol (Agilent, Santa Clara, CA). Concentrations of 1 μM oligomycin, 2 μM FCCP, and 0.5 μM rotenone and antimycin were used (Agilent, Santa Clara, CA). Kinetic stimulation assays were conducted with non-stimulated BMDCs seeded at 2.5 × 105 cells per well in 24 well seahorse plates coated with Cell-Tak (Corning, Corning NY). Stimulants were injected at the concentrations outlined in the APC stimulation section, after the third baseline measurement interval. Metabolic phenotyping was conducted on a Seahorse XFe24 (Agilent, Santa Clara, CA).
Darling R.J., Senapati S., Kelly S.M., Kohut M.L., Narasimhan B, & Wannemuehler M.J. (2019). STING pathway stimulation results in a differentially activated innate immune phenotype associated with low nitric oxide and enhanced antibody titers in young and aged mice. Vaccine, 37(20), 2721-2730.
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7

Coating Glass and Mica with Cell-Tak

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Cell-Tak glass slides were prepared by pipetting 285 ml 100 mM NaHCO3 (pH 8) onto high precision cover glass (24x24 mm2) then 10 μl of Cell-Tak (Corning, 5% (w/v) in acetic acid) and 5 μl of 1 M NaOH. This was covered and left for 20 minutes. The slide was then washed five times with HPLC grade water. The same process with proportionately smaller volumes was used for coating freshly cleaved mica.
Pasquina-Lemonche L., Burns J., Turner R.D., Kumar S., Tank R., Mullin N., Wilson J.S., Chakrabarti B., Bullough P.A., Foster S.J, & Hobbs J.K. (2020). The Architecture of the Gram Positive Bacterial Cell Wall. Nature, 582(7811), 294-297.
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8

Extracellular Acidification Rate in RBC

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The ECAR was measured in a Seahorse XFe24 analyzer as previously reported with slight modifications [15 (link)]. For RBC samples, 5 × 106 cells per well were plated onto XF24 cell plates coated with Cell-Tak (354242, Corning, New York, USA) in XF RPMI assay medium pH 7.4 (100 μL, XF assay medium) supplemented with glucose (40 mM), sodium pyruvate (1 mM) and L-glutamine (2 mM) and left for 10 min at room temperature. After seeding, the cell plates were centrifuged for 1.5 min at 200 × g at room temperature with an acceleration of one without braking. Afterward, 400 μL of XF assay medium was added to the cells, and the cells were incubated for 30 min at 37 °C without CO2. XF measurements included serial injections of glucose (10 mM), oligomycin A (5 μM) and 2-deoxy-glucose (2-DG, 50 mM).
Wu X., Liu Z., Hao D., Zhao Q., Li W., Xie M., Feng X., Liao X., Chen S., Wang S., Zhou C., Long W., Zhong Y., Li S., Cao Y., Wang H., Wang A., Xu Y., Huang M., Liu J., Zhong R., Wu Y, & He Z. (2023). Tyrosine phosphorylation of band 3 impairs the storage quality of suspended red blood cells in the Tibetan high-altitude polycythemia population. Journal of Translational Medicine, 21, 676.
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9

Metabolic Profiling of Alveolar Macrophages

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Cx3cr1CreERT2-IRES-YFP/+Rosa26floxed-tdTomato/+ and CX3CR1-cre mice (control group) received tamoxifen at one day before and one day after Nb inoculation (−1, +1 day) and were sacrificed at day 7. Lung single cells were isolated and td tomato+ and td tomato-alveolar macrophages from reporters or alveolar macrophages from control group mice were sorted via flow cytometry. Alveolar macrophages (~75,000 cells/well) from each group were seeded into CellTak (Corning, # 354240) coated XFp plates in XF RPMI complete assay media, and incubated at 37° C without CO2 for 45 minutes. The assay was run on a Seahorse XFp Extracellular Flux Analyzer with each group of AMs run in duplicate. After 20 cycles of basal measurement, Rotenone and Antimycin A (Seahorse XF Cell Mito Stress Kit), inhibitors of mitochondrial complex I and III, respectively, were injected into each well for a final concentration of 0.5 uM/well. Oxidative phosphorylation was measured by the oxygen consumption rate (OCAR; pmol/min) while glycolysis was measured by the extracellular acidification rate (ECAR; mpH/min) due to the accumulation of lactate or other metabolic acids and release of protons into the extracellular medium.
Chen F., El-Naccache D.W., Ponessa J.J., Lemenze A., Espinosa V., Wu W., Lothstein K., Jin L., Antao O., Weinstein J.S., Damani-Yokota P., Khanna K., Murray P.J., Rivera A., Siracusa M.C, & Gause W.C. (2022). Helminth resistance is mediated by differential activation of recruited monocyte-derived alveolar macrophages and arginine depletion. Cell reports, 38(2), 110215.
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10

Metabolic Profiling of Organoids

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Non-embedded organoids were seeded in a XF 96-well spheroid plate (Agilent Technologies, 102905-100) coated with Cell-Tak (Corning, 354240) and analyzed using an XFe96 Extracellular Flux Analyzer (Seahorse Biosciences) at DIV30. Seahorse XF Cell Mito Stress Tests were carried out using sequential injections of Oligomycin (5 µM), FCCP (1 µM), and Rotenone plus Antimycin A (1 µM). Values were normalized to the size (area) of the organoids. Three baseline measures and three measurements after each compound injection were performed.
Rosety I., Zagare A., Saraiva C., Nickels S., Antony P., Almeida C., Glaab E., Halder R., Velychko S., Rauen T., Schöler H.R., Bolognin S., Sauter T., Jarazo J., Krüger R, & Schwamborn J.C. (2023). Impaired neuron differentiation in GBA-associated Parkinson’s disease is linked to cell cycle defects in organoids. NPJ Parkinson's Disease, 9, 166.
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