Hema 3 stain system

Manufactured by Thermo Fisher Scientific

The Hema-3 Stain System is a laboratory equipment designed for cell staining. It provides a standardized staining process for rapid and consistent results. The system includes staining solutions and an automated staining apparatus to facilitate efficient sample preparation for microscopic analysis.

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Lab products found in correlation

Matrigel basement membrane, by BD (3 mentions) Transwell insert, by Thermo Fisher Scientific (2 mentions)

Close spelling variants of this product

HEMA 3 stain Hema 3 System Hema 3

3 protocols using hema 3 stain system

Assessing Cell Invasiveness via Transwell

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The invasiveness of HeyA8, HeyA8-MDR, and SKOV3IP1 cells was determined as previously described [83 (link)]. Briefly, cells were treated with 50 nM of the indicated oligonucleotide, and 48 hours later, live cells were collected and washed with serum-free media. Fixed numbers of the viable cells (4 × 10⁵ cells), were resuspended in 1 mL of serum-free RPMI-1640 medium and added onto 6-well plate Transwell inserts (8-μm-pore size; Fisher Scientific) coated with a Matrigel basement membrane (0.7 mg/mL; BD Biosciences). Lower chambers were filled with 2 mL of medium supplemented with 15% FBS. After 24 hours, non-invading cells on the upper surface of the filter were removed with cotton swabs. Cells that invaded through the Matrigel onto the lower side of the filter were fixed, stained with the Hema-3 Stain System (Fisher Scientific), and photographed. The number of cells that invaded the lower side of the filter was counted in 5 fields and expressed as the mean number of cells from triplicate measurements.

Rashed M.H., Kanlikilicer P., Rodriguez-Aguayo C., Pichler M., Bayraktar R., Bayraktar E., Ivan C., Filant J., Silva A., Aslan B., Denizli M., Mitra R., Ozpolat B., Calin G.A., Sood A.K., Abd-Ellah M.F., Helal G.K, & Berestein G.L. (2017). Exosomal miR-940 maintains SRC-mediated oncogenic activity in cancer cells: a possible role for exosomal disposal of tumor suppressor miRNAs. Oncotarget, 8(12), 20145-20164.

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Transwell Invasion Assay for Metastatic Potential

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Transwell inserts (4 μm pore size; Fisher Scientific) coated with a Matrigel basement membrane (0.7 mg/mL; BD Biosciences) were used for this assay. After 48-h transfection with 100 nM miR-873, control miRNA, KRAS siRNA, or control siRNA, 8 × 10⁴ (PANC1, MDA-MB-231, and MDA-MB-436) and 100 × 10⁴ (MiaPaCa-2) cells were resuspended in 500 μL serum-free medium and then added to the upper chamber of the insert. Lower chambers were supplied with 500 μL medium supplemented with 10% FBS. After 24 h of incubation, non-invading cells on the upper chamber of the filter were cleared with cotton swabs. Cells that invaded through the Matrigel onto the lower chamber were fixed, stained with the Hema-3 Stain System (Fisher Scientific), and photographed. All experiments were performed in triplicate and invaded cells to the lower side of the filter were counted in at least 5 fields and expressed as a percentage of invasion.

Mokhlis H.A., Bayraktar R., Kabil N.N., Caner A., Kahraman N., Rodriguez-Aguayo C., Zambalde E.P., Sheng J., Karagoz K., Kanlikilicer P., Abdel Aziz A.A., Abdelghany T.M., Ashour A.A., Wong S., Gatza M.L., Calin G.A., Lopez-Berestein G, & Ozpolat B. (2018). The Modulatory Role of MicroRNA-873 in the Progression of KRAS-Driven Cancers. Molecular Therapy. Nucleic Acids, 14, 301-317.

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Invasion Assay for ZA Treatment

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The invasiveness of HeyA8-MDR and OVCAR-5 cells treated with ZA was determined as previously described (15 (link)). Briefly, HeyA8-MDR and OVCAR-5 cells were collected and washed with serum-free media. Cells (4×10⁵) were resuspended in 1mL of serum-free DMEM/F-12 medium and added onto 6-well plate transwell inserts (8µm pore-size; Fisher Scientific, Middleton, VA, USA) coated with a Matrigel basement membrane (0.7mg/mL; BD Biosciences, Bedford, MA, USA). Lower chambers were filled with 2mL of DMEM/F-12 medium supplemented with 10% FBS. HeyA8-MDR and OVCAR-5 cells in the transwell inserts were then treated with different concentrations of ZA in serum-free media. Seventy-two hours later, non-invading cells on the upper surface of the filter were removed with cotton swabs. Cells that invaded through the Matrigel onto the lower side of the filter were fixed, stained with the Hema-3 Stain System (Fisher Scientific), and photographed. For each filter, the number of invaded cells was counted in nine fields, and expressed as the mean number of cells from triplicate measurements.

Gonzalez-Villasana V., Fuentes-Mattei E., Ivan C., Dalton H.J., Rodriguez-Aguayo C., Fernandez-de Thomas R.J., Aslan B., Monroig P.D., Velazquez-Torres G., Previs R.A., Pradeep S., Kahraman N., Wang H., Kanlikilicer P., Ozpolat B., Calin G., Sood A.K, & Lopez-Berestein G. (2015). Rac1/Pak1/p38/MMP-2 axis regulates angiogenesis in ovarian cancer. Clinical cancer research : an official journal of the American Association for Cancer Research, 21(9), 2127-2137.

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