Cell Meter™ Cell Adhesion Assay Kit

The Cell Meter™ Cell Adhesion Assay Kit is a fast and sensitive assay for measuring cell-cell or cell-surface adhesion for a variety of cell types. In this assay, cells are labeled with Calcein UltraGreen AM and allowed to adhere. After removal of nonadherent cells, The fluorescence of Calcein UltraGreen is used to calculate the number of adherent cells. The use of our outstanding fluorogenic dye, Calcein UltraGreen AM provides a fast and sensitive method for measuring cell adhesion with a variety of cell types. Calcein UltraGreen AM is nonfluorescent but, once loaded into cells, is cleaved by endogenous esterases to produce highly fluorescent Calcein UltraGreen, a brightly fluorescent, pH-independent, cytoplasmic cell marker with the minimal interference to cell adhesion process. The Cell Meter™ cell adhesion assay is designed for use with fluorescence microplate readers. The robust performance of Calcein UltraGreen AM and simple procedure of the kit avoids problems associated with assays that utilize radioisotopes, which generate hazardous waste, and with assays that rely on the use of covalently coupled cell-surface labels, which can potentially alter cell function.

Example protocol

AT A GLANCE

Protocol summary

Add cells on a plate coated with desired coating material
Incubate cells at 37 °C to allow them to attach
Remove the unattached cells
Add Calcein Ultragreen AM working solution
Incubate the cells at 37 °C for 20-30 minutes
Remove supernatant and wash cells with HHBS or DPBS
Measure the fluorescence intensity using fluorescence microplate reader with Ex/Em = 490/525 nm

Important

Thaw all the kit components at room temperature before starting the experiment.

PREPARATION OF STOCK SOLUTIONS

Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.

Calcein Ultragreen AM stock solution

Add 50 µL of DMSO (Component C) into Calcein Ultragreen AM (Component A) and mix well.
Note Store the unused Calcein Ultragreen AM stock solution at -20 °C in single use aliquots to avoid freeze thaw cycles.

PREPARATION OF WORKING SOLUTION

Calcein Ultragreen AM working solution

Add 50 µL of Calcein Ultragreen AM stock solution into 10 mL of Adhesion Assay Buffer and mix well.
Note Calcein Ultragreen AM working solution should not be stored and should be used promptly.
Note 10 mL Calcein Ultragreen AM working solution is enough for 100 tests.

SAMPLE EXPERIMENTAL PROTOCOL

The following protocol can be used as a guideline and should be optimized according to the needs.

Add 100 µL volumes of cells on a plate coated with desired coating material.
Incubate plate at 37 °C for 2 to 3 hours.
Note For each cell line, optimal incubation time should be tested experimentally.
Remove the growth medium and unattached cells.
Add 100 µL of Calcein Ultragreen AM working solution and incubate plate at 37 °C for 20-30 minutes.
Note For each cell line, optimal incubation time should be tested experimentally.
Remove the dye working solution and wash cells with 1X Hank’s salt solution and 20 mM Hepes buffer or DPBS once.
Add 100 µL of Adhesion Assay Buffer to the wells.
Monitor the fluorescence intensity using a fluorescence microplate reader at Ex/Em = 490/525 nm (Cutoff = 515 nm).

Spectrum

Open in Advanced Spectrum Viewer

Product family

Name	Excitation (nm)	Emission (nm)
Cell Meter™ Cell Viability Assay Kit Blue Fluorescence with 405 nm Excitation	406	445
Cell Meter™ Cell Viability Assay Kit Green Fluorescence	494	514
Cell Meter™ Cell Proliferation Assay Kit	503	527

Citations

View all 1 citations: Citation Explorer

Hpgd affects the progression of hypoxic pulmonary hypertension by regulating vascular remodeling
Authors: He, Meng and Tao, Kelong and Xiang, Min and Sun, Jian
Journal: BMC Pulmonary Medicine (2023): 1--12

References

View all 50 references: Citation Explorer

The Protein A-mediated binding of Staphylococcus to antibodies in flow cytometric assays and its reduction using FcR blocking reagent.
Authors: Cronin, Ultan P and Girardeaux, Laura and O'Meara, Elaine and Wilkinson, Martin
Journal: Applied and environmental microbiology (2020)

Effect of advanced glycation end product on paraoxonase 2 expression: Its impact on endoplasmic reticulum stress and inflammation in HUVECs.
Authors: Ravi, Ramya and Ragavachetty Nagaraj, Nareshkumar and Subramaniam Rajesh, Bharathidevi
Journal: Life sciences (2020): 117397

Functionalized Graphene Nanoparticles Induce Human Mesenchymal Stem Cells to Express Distinct Extracellular Matrix Proteins Mediating Osteogenesis.
Authors: Newby, Steven D and Masi, Tom and Griffin, Christopher D and King, William J and Chipman, Anna and Stephenson, Stacy and Anderson, David E and Biris, Alexandru S and Bourdo, Shawn E and Dhar, Madhu
Journal: International journal of nanomedicine (2020): 2501-2513

Passaging capability of human corneal endothelial cells derived from old donors with and without accelerating cell attachment.
Authors: Parekh, Mohit and Peh, Gary and Mehta, Jodhbir S and Ramos, Tiago and Ponzin, Diego and Ahmad, Sajjad and Ferrari, Stefano
Journal: Experimental eye research (2019): 107814

CD56 expression in breast cancer induces sensitivity to natural killer-mediated cytotoxicity by enhancing the formation of cytotoxic immunological synapse.
Authors: Taouk, Ghina and Hussein, Ola and Zekak, Moussa and Abouelghar, Ali and Al-Sarraj, Yasser and Abdelalim, Essam M and Karam, Manale
Journal: Scientific reports (2019): 8756

Page updated on November 20, 2024

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