Ultra low attachment 96 well u bottom plates

Manufactured by Corning

Sourced in United States

The Corning Ultra-low-attachment 96-well U-bottom plates provide a specialized surface for cell culture applications that minimizes cell attachment. These plates are designed to promote the formation of cellular aggregates, spheroids, and suspension cultures.

Automatically generated - may contain errors

Lab products found in correlation

Anti mouse f4 80 antibody, by BD (2 mentions) Mtesr medium, by STEMCELL (1 mentions) Anti human cd45 antibody, by BioLegend (1 mentions) Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions) Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (1 mentions) Anti cd47 clone b6h12, by BioXCell (1 mentions) Tryple express, by Thermo Fisher Scientific (1 mentions) Guava easycyte, by Merck Group (1 mentions) Anti human cd45, by BD (1 mentions) Fortessa x20 flow cytometer, by BD (1 mentions) Interleukin 3 (il 3), by Thermo Fisher Scientific (1 mentions) X vivo 15 media, by Lonza (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Glutamax, by Thermo Fisher Scientific (1 mentions) M csf, by Thermo Fisher Scientific (1 mentions) Mtesr media, by STEMCELL (1 mentions)

Close spelling variants of this product

Ultra-low attachment U-bottom 96-well plate Ultra-low cell attachment U-bottom 96-well plates Costar Ultra low attachment U-bottom 96-well plates Low attachment 96-well U-bottom plate U-bottom ultra-low attachment plates Ultra-low attachment 96-well plates Well ultra-low attachment plates Low-attachment 96-well plates 96-well U-bottom plates U-bottom 96 wells plate

8 protocols using ultra low attachment 96 well u bottom plates

Phagocytosis Assay of Human and Mouse Macrophages

Check if the same lab product or an alternative is used in the 5 most similar protocols

For the phagocytosis assay of human primary macrophage, A2780 cells prelabeled with CFSE (Thermo Fisher, C34554) were cocultured with human macrophages at a ratio of 2:1 for 4 hours in the presence of vehicle control, αCD47-G1 or αCD47-G4 at the dose of 5 μg/ml or supernatants of OV-infected cells at 37 °C in ultra-low-attachment 96-well U-bottom plates (Corning) in serum-free 1640 (Life Technologies). Then the cells were harvested and stained with anti-human CD45 (BD, 552850) to identify macrophages. All flow cytometry data were collected using a Fortessa X20 flow cytometer (BD Biosciences). Phagocytosis was calculated as the number of CD45⁺CFSE⁺ macrophages, quantified as a percentage of the total CD45⁺ macrophages.
For the phagocytosis of mouse macrophages, ID8 or A2780 cells prelabeled with CFSE were cocultured with murine macrophages at a ratio of 2:1 for 6 hours with 5 μg/ml antibodies or supernatants of OV-infected cells at 37°C in ultra-low-attachment 96-well U-bottom plates (Corning) in the serum-free 1640 media (Life Technologies). The cells were harvested. An anti-mouse F4/80 antibody (BD, cat#565787) was used to identify macrophages. Phagocytosis was measured as the number of F4/80⁺CFSE⁺ macrophages and quantified as a percentage of the total F4/80⁺ macrophages.

Tian L., Xu B., Teng K.Y., Song M., Zhu Z., Chen Y., Wang J., Zhang J., Feng M., Kaur B., Rodriguez L., Caligiuri M.A, & Yu J. (2021). Targeting Fc receptor-mediated effects and the “don’t eat me” signal with an oncolytic virus expressing an anti-CD47 antibody to treat metastatic ovarian cancer. Clinical cancer research : an official journal of the American Association for Cancer Research, 28(1), 201-214.

+ Open protocol

+ Expand

Tumor Sphere Formation Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

For the tumor sphere formation assay, 1 × 10³ Caki-1 or 786-O cells were seeded into U-bottom ultra-low attachment 96-well plates (Corning, cat. no. 174925) and cultured in DMEM supplemented with 10% FBS for 2 weeks. Tumor sphere images were captured under a microscope and the area was measured with Image J software (National Institutes of Health).

Ma W., Li X., Yang L., Pan J., Chen Y., Lu Y., Dong X., Li D, & Gan W. (2022). High VSX1 expression promotes the aggressiveness of clear cell renal cell carcinoma by transcriptionally regulating FKBP10. Journal of Translational Medicine, 20, 554.

+ Open protocol

+ Expand

Fluorescent JEG-3 Trophoblast Spheroids

Check if the same lab product or an alternative is used in the 5 most similar protocols

Fluorescent spheroids of the trophoblast cell line JEG-3 were generated by transduction with recombinant lentiviral particles expressing GFP from a PGK promoter (Tiscornia et al., 2006) . Briefly, trophoblast cells were grown on 12well plates until cultures reached 70% confluence. Lentiviral particles were added to the culture at a multiplicity of infection (MOI) of 10. Trophoblast cells with the highest GFP levels were collected by fluorescence-activated cell sorting (FACS) and cultured further. To generate spheroids, suspensions of GFP+ JEG-3 cells were adjusted to a 3 × 10 4 cells/ml concentration; 100 µl were seeded in U-bottom ultra-low attachment 96-well plates (Corning, NY, USA) and centrifuged at room temperature for 10 min at 250g. The plates were kept for 48 h at 37°C and 5% CO 2 , resulting in spheroids of around 250 µm diameter.

Vergaro P., Tiscornia G., Zambelli F., Rodríguez A., Santaló J, & Vassena R. (2021). Trophoblast attachment to the endometrial epithelium elicits compartment-specific transcriptional waves in an in-vitro model. Reproductive biomedicine online, 42(1).

+ Open protocol

+ Expand

Cerebral Organoid Generation from Human Embryonic Stem Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human embryonic stem cells (hESCs) were obtained from WiCell, and cultured in a feeder‐free condition. Cells were maintained with mTeSR medium (Stemcell Technologies) on the Matrigel‐coated 6‐well plates at 37°C supplied with 5% CO₂. Cells were cultured and passaged using standard procedures according to the previous description.²⁴ Normal karyotype and contamination‐free were confirmed.
Cerebral organoids were cultured as a previous publication²⁵ with slight modifications. Briefly, H9 hESCs were treated with 0.5 mmol/L EDTA and Accutase to obtain single‐cell suspension. Embryoid bodies (EBs) were generated with 9000 cells/well in the U‐bottom, Ultra low‐attachment 96‐well plates (Corning) with 150 µL of mTeSR medium containing 1xRevitaCell supplement (Gibco) at day 0. Fresh mTeSR medium without RevitaCell supplement was fed to EBs at day 3. At day 5, EBs were transferred into Neural Induction (NI) medium, and medium was exchanged with fresh NI medium every second day for 6 days. EBs were then embedded into Matrigel droplets, and cultured in differentiation medium without vitamin A and shaking. Five days later, cerebral organoids were cultured in differentiation medium supplied with vitamin A on an orbital shaker. Media were exchanged every 5 days until day 30, and used for further experiments.

Jiang M., Tang T., Liang X., Li J., Qiu Y., Liu S., Bian S., Xie Y., Fang F, & Cang J. (2021). Maternal sevoflurane exposure induces temporary defects in interkinetic nuclear migration of radial glial progenitors in the fetal cerebral cortex through the Notch signalling pathway. Cell Proliferation, 54(6), e13042.

+ Open protocol

+ Expand

Macrophage Phagocytic Activity Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

Primary human or mouse macrophages were incubated with carboxyfluorescein succinimidyl ester (CFSE)-labeled (Thermo Fisher Scientific, catalog no. C34554) GBM30 or CT2A-hEGFR target cells, respectively, at an effector/target ratio of 1:2 for 6 h in the presence 5 μg ml⁻¹ or 10 μg ml⁻¹ Cmab-h(m)CCL5 fusion protein or IgG1 isotype control in a humidified, 5% CO₂ incubator at 37 °C in ultra-low-attachment 96-well U-bottom plates (Corning) in serum-free 1640 media (Life Technologies). The co-cultured cells were harvested by centrifuging them at 400 × g for 5 min at 4 °C and stained with an anti-human CD45 antibody (BioLegend, catalog no. 368516) or an anti-mouse F4/80 antibody (BD Biosciences, catalog no. 565787) to identify human or mouse macrophages, respectively, in flow cytometry assay. Human macrophage phagocytosis was quantified as the percentage of CD45⁺CFSE⁺ macrophages among total CD45⁺ cells. Murine macrophage phagocytosis was quantified as the percentage of F4/80⁺CFSE⁺ macrophages among total F4/80⁺ macrophages. The gating strategy is shown in Supplementary Fig. 2.

Zhu Q.Y., Headrick J.P, & Berne R.M. (1991). Transmural distribution of extracellular purines in isolated guinea pig heart.. Proceedings of the National Academy of Sciences of the United States of America, 88(2), 657-660.

+ Open protocol

+ Expand

In Vitro Phagocytosis Assay for Tumor Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

For all flow-based in vitro phagocytosis assays, tumor cells and human macrophages were co-cultured at a ratio of 2:1 in ultra-low-attachment 96-well U-bottom plates (Corning) in serum-free RPMI (Thermo Fisher Scientific). Tumor cells either virally expressed GFP or were labeled with CFSE (Invitrogen) by suspending cells in PBS (5 μM working solution) as per manufacturer instructions for 20 min at 37 °C protected from light and washed twice with 20 ml of FBS-containing media before co-culture. Anti-GD2 (dinutuximab, acquired from United Therapeutics) or anti-B7-H3 (enoblituzumab, acquired from MacroGenics) was added alone or in combination with anti-CD47 (clone B6H12, Bio X Cell) at a concentration of 10 μg ml⁻¹ (or the concentration indicated in the figures). Tumor cells and antibodies were incubated for 30 min in a humidified 5% CO₂ incubator at 37 °C. Plates were washed two times; human macrophages were added to the plate; and plates were incubated for 1–2 h at 37 °C. Phagocytosis was stopped by washing with 4 °C PBS and centrifugation at 336g before the cells were stained with Live/Dead stain and anti-CD11b. Assays were analyzed by flow cytometry, and phagocytosis was measured as the number of CD11b⁺ and CFSE/GFP⁺ macrophages, quantified as a percentage of the total CD11b⁺ macrophages and normalized to the control condition.

Theruvath J., Menard M., Smith B.A., Linde M.H., Coles G.L., Dalton G.N., Wu W., Kiru L., Delaidelli A., Sotillo E., Silberstein J.L., Geraghty A.C., Banuelos A., Radosevich M.T., Dhingra S., Heitzeneder S., Tousley A., Lattin J., Xu P., Huang J., Nasholm N., He A., Kuo T.C., Sangalang E.R., Pons J., Barkal A., Brewer R.E., Marjon K.D., Vilches-Moure J.G., Marshall P.L., Fernandes R., Monje M., Cochran J.R., Sorensen P.H., Daldrup-Link H.E., Weissman I.L., Sage J., Majeti R., Bertozzi C.R., Weiss W.A., Mackall C.L, & Majzner R.G. (2022). Anti-GD2 synergizes with CD47 blockade to mediate tumor eradication. Nature medicine, 28(2), 333-344.

+ Open protocol

+ Expand

Tumor Cell Phagocytosis by Macrophages

Check if the same lab product or an alternative is used in the 5 most similar protocols

All in vitro phagocytosis assays reported here were performed by co-culturing tumor cells and macrophages at a ratio of 200,000 tumor cells to 100,000 macrophages for 2 h in a humidified, 5% CO₂ incubator at 37 °C in ultra-low-attachment 96-well U-bottom plates (Corning). Tumor cells expressing fluorescence GFP were collected from plates using TrypLE Express (Life Technologies) before co-culture. After co-culture, phagocytosis assays were stopped. Cells were centrifuged at 400 g for 5 min at 4 °C and stained with APC-labelled anti-CD45 (Clone I3/2.3, BioLegend) to identify macrophages. Assays were analyzed by flow cytometry on Guava easyCyte (Millipore). Phagocytosis was measured as the number of CD45⁺GFP⁺ macrophages, quantified as the percentage of the total CD45⁺macrophages.

Zhang C., Liu Y., Jiang J., Chen C., Duan Z., Su H., Wang S., Tian B., Shi Y., Xiang R, & Luo Y. (2024). Targeting tumor cell-to-macrophage communication by blocking Vtn-C1qbp interaction inhibits tumor progression via enhancing macrophage phagocytosis. Theranostics, 14(7), 2757-2776.

+ Open protocol

+ Expand

Generating Human iPSC-Derived Macrophages

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human iPSCs derived from umbilical cord blood CD34 + cells (15, (46) (47) (48) were maintained as undifferentiated cells on matrigel-coated tissue culture flasks in mTeSR media (STEMCELL Technologies). Human iPSC-derived macrophages were generated according to previously described protocol (49) with some modifications. Briefly, 8000 human iPSCs were single cell passaged and then aggregated into spin embryoid bodies (EBs) by centrifugation on ultra-low attachment 96-well u-bottom plates (Corning) with cytokines that are essential for hematopoietic progenitor development (APEL media containing 40 ng/mL SCF, 20 ng/mL VEGF, 20 ng/mL BMP4 and 10 μM Rock-Y).
EBs were then manually transferred (20 EBs per well) onto 0.1% gelatin-coated 6-well plates containing differentiation medium I (X-VIVO15 media (Lonza) supplemented with 1% penicillin-streptomycin (Gibco), 1% GlutaMAX (Gibco), 55 mM 2-Mercapnoethanol (Gibco), 50 ng/mL M-CSF and 25 ng/mL IL-3 (all from Peprotech)). Human iPSC-derived macrophage progenitor cells (iPSC-MPro) started to generate after 1-2 weeks. These cells were then collected and further differentiated to mature macrophages (human iPSC-derived macrophages) in differentiation medium II (X-VIVO15 media supplemented with 1% penicillin-streptomycin, 1% GlutaMAX and 100 ng/mL M-CSF) for 7 days.

Pouyanfard S., Meshgin N., Cruz L.S., Diggle K., Hashemi H., Pham T.V., Fierro M., Tamayo P., Fanjul A., Kisseleva T, & Kaufman D.S. (2021). Human induced pluripotent stem cell-derived macrophages ameliorate liver fibrosis. Stem cells (Dayton, Ohio), 39(12).

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!