DNA Flow Cytometry Reagents

Cell Viability Stain: One step, and ready-to-use. Allows for the rapid staining of dead cells by membrane penetration using propidium iodide.

DNA Standard for Flow Cytometry: Trout Red Blood Cell (TRBC) based DNA standard that is non-toxic and optimized for flow cytometry.

Nuclear Isolation Medium (NIM): Isolates intact nuclei from cells, tissues, frozen-thawed tissues, or deparafinized/enzymatically isolated tissues.

From the laboratory of Jerry T. Thornthwaite, PhD, Cancer Research Institute of West Tennessee.

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Catalog Number Product Size AVAILABILITY Price Qty
ECT001
Cell Viability Stain, 10mL (100 reactions)
10mL (100 reactions) 2-3 weeks
Regular Price:$65.00
ECT002
DNA Standard for Flow Cytometry, 5mL (100 reactions)
5mL (100 reactions) In stock
Regular Price:$53.00
ECT003
Nuclear Isolation Medium, 100mL (100 reactions)
100mL (100 reactions) In stock
Regular Price:$108.00
ECT004
Nuclear Isolation Medium-DAPI, 100mL (100 reactions)
100mL (100 reactions) 2-3 weeks
Regular Price:$134.00
ECT005
Nuclear Isolation Medium-Propidium Iodide, 100mL (100 reactions)
100mL (100 reactions) In stock
Regular Price:$134.00
Specifications

Product Type: Buffer or Chemical
Name: Cell Viability StainDNA Standard for Flow CytometryNuclear Isolation Medium (NIM)
Amount: Cell Viability Stain: 10mL (100 reactions)DNA Standard for Flow Cytometry: 5mL of 1-2 x 106 TRBC/mL (100 reactions)Nuclear Isolation Medium (NIM): 100mL (100 reactions)
Storage: 4C
Shipped: Cold packs

Documentation

Suggested Protocol

Cell Viability Stain: Mix 100uL of Cell Viability Stain with 100uL of cells at 1x10^5-1x10^6 cells/mL)

DNA Standard for Flow Cytometry: Add 50uL of the DNA Standard to 1mL of cells at 1-2x10^6 cells/mL. Centrifuge to a pellet and resuspend in either NIM only (customized staining), NIM-DAPI, or NIM-PI. Alternatively, one may add portions of the DNA standard and cells directly to 1 mL of the NIM solutions and stain without centrifugation. The suspensions should be kept on ice or room temperature for 2-3 minutes before analyzing.

Nuclear Isolation Medium (NIM): In conjunction with the DNA Standard (see above), resuspend centrifuged pellet in 100uL of either NIM only (for customized staining), NIM-DAPI, or NIM-PI. Alternatively, one may add portions of the DNA standard and cells directly to 1 mL of the NIM solutions and stain without centrifugation. The suspensions should be kept on ice or room temperature for 2-3 minutes before analyzing.

Data

Tonsil with TRBC DNA Standard and Nuclear Isolation Medium (NIM-DAPI)

The DNA Standard was mixed with NIM-DAPI, and filtered through a 37µm filter. The DNA standard served to establish the flow cytometer was operating within the precise boundaries of CV= 1-2%. Secondly, during a sample run, the DNA standard peak at channel 50 (approximately 5.0 pg/ nucleus). The position in channel 50 was used to assure the para enzymes were functioning by using the human tonsil to determine the optimum CV= 2-3% and the relative DNA value for the G0 tonsil nuclei of 7.4 pg/nucleus. Control paraffinated human tonsil tissues were also subjected to deparaffination and enzymatic dissociation to assure the PARA reagents were functioning properly. All tonsil controls were costhin a CV= 2-3% for each determination of DNA histogram measurements of the benign and cancer tissues. Also, the CV= 1-2% for all DNA standard samples. (A) Typical DNA Standard Trout Red Blood Cell (TRBC) Values. CV = 1.42 ± 0.19 SD, n=66. (B) Human Tonsil with DNA Standard. The data for 22 samples resulted in an average coefficient of variation of 2.18 ± 0.46 SD, an average DNA index of 1.42 ± 0.03 SD, and an average S-phase fraction of 4.37 ± 1.37 SD.

Adapted from: Thornthwaite, J.T., et al. J. Solid Tumors 3: 12-23 (2013).

Provider
From the laboratory of Jerry T. Thornthwaite, PhD, Cancer Research Institute of West Tennessee.
References
  1. Thornthwaite, J.T., Stanfill, J.C., Ahmed, A.S. Comparison of clinical staging of benign and malignant ovarian tissues with DNA flow cytometry. J. Solid Tumors 3: 12-23 (2013).
  2. Thornthwaite, J.T., et al. Anticancer Effects of Curcumin, Artemisinin, Genistein, and Resveratrol, and Vitamin C: Free Versus Liposomal Forms. Advances in Biological Chemistry, 7, 27-41 (2017).

If you publish research with this product, please let us know so we can cite your paper.

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