Cell based assay fixative solution

Manufactured by Cayman Chemical

Cell-based assay fixative solution is a laboratory product designed to preserve the structure and morphology of cells during the sample preparation process for various cell-based assays. It is a ready-to-use solution that can be applied to cell cultures to fix and stabilize the cells, enabling effective analysis and quantification.

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Lab products found in correlation

Ldl dylight 550, by Cayman Chemical (1 mentions) Tcs sp8 confocal microscope, by Leica (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Cholesterol cell based detection assay kit, by Cayman Chemical (1 mentions) Dm il led microscope, by Leica (1 mentions)

2 protocols using cell based assay fixative solution

LDL Uptake and Receptor Assay

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For LDL uptake cell-based assay, the cells were treated with LDL-DyLight^TM 550 (Cayman chemical, Ann Arbor, Michigan, USA) working solution in serum-free medium. Incubate the cells at 37 °C with 5% CO₂ for an additional 18 hours. At the end of LDL uptake incubation, the culture medium was replaced with fresh culture medium. The degree of LDL uptake was examined under a Leica TCS SP8 confocal microscope (Leica, Wetzlar, Germany) with filters capable of measuring excitation and emission wavelengths 540 and 570 nm, respectively.
For LDL receptors examination, the cells were washed with TBS briefly, and fixed with cell-based assay fixative solution (Cayman chemical) for 10 minutes. Wash the cells with TBST three times for five minutes each. Incubate the cells for one hour with rabbit anti-LDL receptor antibody (Cayman chemical). Wash the cells with TBST three times for five minutes each. Incubate the cells for one hour with Dylight^TM 488-conjugated secondry antibody (Cayman chemical). Wash the cells with TBST three times for five minutes each. The nuclei were counter-stained with DAPI (Sigma-Aldrich). The stained cells were examined under a Leica TCS SP8 confocal microscope (Leica, Wetzlar, Germany).

Huang J., Guo X., Li W, & Zhang H. (2017). Activation of Wnt/β-catenin signalling via GSK3 inhibitors direct differentiation of human adipose stem cells into functional hepatocytes. Scientific Reports, 7, 40716.

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Modulating Viral Infectivity via Cholesterol

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PAM-pCD163 and Vero cells were first preincubated with vehicle or MβCD at various final concentrations for 1 h and then supplemented with or without 100 μg/ml or 40 μg/ml exogenous cholesterol, respectively, and incubated for 1 h. The cells were then inoculated with PRRSV or PEDV as described above. The virus inoculum was removed, and the infected cells were maintained in fresh medium containing MβCD and exogenous cholesterol. At 48 h dpi, the virus-infected cells were harvested and subjected to FACS analysis to assess the infectivity of PRRSV and PEDV as described above. In parallel, the cellular cholesterol content was determined using a Cholesterol Cell-Based Detection Assay Kit (Cayman Chemical) according to the manufacturer’s instructions. Briefly, virus-infected cells were cultivated in the presence of MβCD and exogenous cholesterol for 48 h, fixed with Cell-Based Assay Fixative Solution (Cayman Chemical) for 10 min at RT, and then washed three times for 5 min each with Cell-Based Assay Wash Buffer (Cayman Chemical). The cells were incubated with filipin III for 1 h in the dark. After washing three times in wash buffer, the cells were counterstained with DAPI, and filipin staining was visualized using a fluorescent Leica DM IL LED microscope.

Jeon J.H, & Lee C. (2017). Cellular cholesterol is required for porcine nidovirus infection. Archives of Virology, 162(12), 3753-3767.

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