Tc252

Manufactured by HiMedia

The TC252 is a centrifuge designed for general laboratory use. It has a maximum rotor speed of 6,000 RPM and a maximum RCF of 4,000 x g. The unit features a sturdy construction and includes a safety interlock system.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions) Mb031, by HiMedia (1 mentions) Alexafluortm 633 conjugated phalloidin, by Thermo Fisher Scientific (1 mentions) Ix73 fluorescence microscope, by Olympus (1 mentions) Calcein am, by Thermo Fisher Scientific (1 mentions) Propidium iodide, by HiMedia (1 mentions)

3 protocols using tc252

3D Spheroid Imaging and Viability Assay

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Spheroids were first pelleted in a sterile 15-mL tube. Cells were then fixed using 3.7% formaldehyde (24005, Thermo Fisher Scientific) at

4^{\circ}

C for 30 min. After fixation, cells were washed and resuspended in PBS. Following this, 20 μL of spheroid suspension was dried in eight well chambered cover glass at

37^{\circ}

C for 30 min. Permeabilization was achieved using 0.5% Triton X-100 in PBS (PBST) (MB031, HiMedia) for 2h at RT.

{Alexa Fluor}^{T M}

633-conjugated phalloidin (A22284, Thermo Fisher Scientific) was added to cells at 1:500 dilution in 0.1% PBST and incubated overnight at

4^{\circ}

C. Cells were washed thrice with 1

\times

PBS for 5 min, counterstained with 1 μg/mL DAPI (D1306, Thermo Fisher Scientific).
To determine live and dead cells in the culture, cells were stained with 0.5 mg/mL Calcein AM (C1430, Invitrogen) for 15 min. Cells were washed with PBS, counterstained with Propidium Iodide (TC252, HiMedia) for 5 min, and imaged under the Olympus IX73 fluorescence microscope.

Arora G., Banerjee M., Langthasa J., Bhat R, & Chatterjee S. (2023). Targeting metabolic fluxes reverts metastatic transitions in ovarian cancer. iScience, 26(11), 108081.

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Quantifying Cell Adhesion on Coated Surfaces

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MDA-MB-231 high and medium α-2,6-sialic
acid cells were trypsinized. After counting, 30 000 cells per
well were incubated in BSA- and ECM-coated wells for 30 min at 37
°C. Unadhered cells were removed carefully, and wells were washed
with 1× phosphate buffered saline pH 7.4 (PBS) thrice to remove
unadhered cells. Cells were fixed using 100% methanol for 10 min at
room temperature. After fixing, cells were washed with 1× PBS
thrice, stained with 50 μg/mL propidium iodide (HiMedia, TC252)
for 30 min at room temperature, and washed thrice with 1× PBS.
Using the plate reader, fluorescence was read at Ex 535 nm/Em 617
nm. BSA or ECM without cells was used as the blank. The assay was
done in triplicates and repeated three times independently.

Pally D., Pramanik D., Hussain S., Verma S., Srinivas A., Kumar R.V., Everest-Dass A, & Bhat R. (2021). Heterogeneity in 2,6-Linked Sialic Acids Potentiates Invasion of Breast Cancer Epithelia. ACS Central Science, 7(1), 110-125.

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Quantifying Cell Adhesion and Proliferation

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MDA-MB-231 high and medium α-2,6 sialic acid cells were trypsinized. After counting, 30,000 cells per well were incubated in BSA and ECM coated wells for 30 min at 37 o C. Unadhered cells were removed carefully and wells were washed with 1X phosphate buffered saline pH 7.4 (PBS) thrice to remove unadhered cells. Cells were fixed using 100% methanol for 10 min at room temperature. After fixing, cells were washed with 1X PBS thrice and stained with 50 μg/mL propidium iodide (HiMedia, TC252) for 30 min at room temperature. Remove excess stain and wash cells thrice with 1x PBS. Using plate reader, fluorescence was read using Ex 535nm/Em 617nm. BSA or ECM without cells was used as blank. Assay was done in triplicates and repeated three times independently.

Pally D., Pramanik D., Hussain S., Verma S., Srinivas A., Pandey A., & Bhat R. (2020). ST6GAL1-mediated heterogeneity in sialic acid levels potentiates invasion of breast cancer epithelia.

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