The Nuclei Extraction Buffer is designed to gently, rapidly, and effectively generate single nuclei suspensions from fresh and frozen tissues. It has been developed for use with the gentleMACS Dissociators and C Tubes.

Data and images for Nuclei Extraction Buffer

Figures

Figure 1

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Preparation of single nuclei suspensions from mouse brain and tumors.
The figure shows the numbers of extracted nuclei per mg of dissociated mouse brain and syngeneic mouse tumors, as indicated in the figure legend.
Single nuclei suspensions were obtained from fresh and frozen tissues samples by using the Nuclei Extraction Buffer in combination with the gentleMACS Octo Dissociator. Nuclei were stained with DAPI and analyzed by flow cytometry using the MACSQuant 10 Analyzer.

Figure 1

Preparation of single nuclei suspensions from mouse brain and tumors.
The figure shows the numbers of extracted nuclei per mg of dissociated mouse brain and syngeneic mouse tumors, as indicated in the figure legend.
Single nuclei suspensions were obtained from fresh and frozen tissues samples by using the Nuclei Extraction Buffer in combination with the gentleMACS Octo Dissociator. Nuclei were stained with DAPI and analyzed by flow cytometry using the MACSQuant 10 Analyzer.

Figure 2

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Preparation of single nuclei suspensions from snap frozen mouse tissue and OCT embedded human tumors.
The figure shows the numbers of extracted nuclei per mg of dissociated snap frozen mouse tissues, including liver, heart and kidney and from OCT embedded human tumors, including melanoma, breast cancer, colorectal cancer and prostatic cancer by using the Nuclei Extraction Buffer in combination with the gentleMACS Octo Dissociator.
Nuclei were stained with DAPI and analyzed by flow cytometry using the MACSQuant 10 Analyzer.

Figure 2

Preparation of single nuclei suspensions from snap frozen mouse tissue and OCT embedded human tumors.
The figure shows the numbers of extracted nuclei per mg of dissociated snap frozen mouse tissues, including liver, heart and kidney and from OCT embedded human tumors, including melanoma, breast cancer, colorectal cancer and prostatic cancer by using the Nuclei Extraction Buffer in combination with the gentleMACS Octo Dissociator.
Nuclei were stained with DAPI and analyzed by flow cytometry using the MACSQuant 10 Analyzer.

Figure 3

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Nuclei extracted with the Nuclei Extraction Buffer maintain RNA integrity.
The table shows the RNA integrity number (RIN) values obtained after RNA isolation from nuclei extracted from snap-frozen and fresh mouse tissue samples using the Nuclei Extraction Buffer in combination with the gentleMACS Octo Dissociator.

Figure 3

Nuclei extracted with the Nuclei Extraction Buffer maintain RNA integrity.
The table shows the RNA integrity number (RIN) values obtained after RNA isolation from nuclei extracted from snap-frozen and fresh mouse tissue samples using the Nuclei Extraction Buffer in combination with the gentleMACS Octo Dissociator.

Figure 4

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Single nuclei suspension from mouse liver.
Single nuclei were extracted from snap-frozen mouse liver tissue using the Nuclei Extraction Buffer in combination with the gentleMACS™ Octo Dissociator. Immediately after nuclei extraction, nuclei were stained using DRAQ5™ Staining Solution. The image shows an overlay of DRAQ5™ (pink) and brightfield.

Figure 4

Single nuclei suspension from mouse liver.
Single nuclei were extracted from snap-frozen mouse liver tissue using the Nuclei Extraction Buffer in combination with the gentleMACS™ Octo Dissociator. Immediately after nuclei extraction, nuclei were stained using DRAQ5™ Staining Solution. The image shows an overlay of DRAQ5™ (pink) and brightfield.

Figure 5

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Single nuclei suspension from mouse brain.
Single nuclei were extracted from snap-frozen mouse brain using the Nuclei Extraction Buffer in combination with the gentleMACS™ Octo Dissociator. Immediately after nuclei extraction, nuclei were stained using DRAQ5™ Staining Solution. The image shows an overlay of DRAQ5™ (pink) and brightfield.

Figure 5

Single nuclei suspension from mouse brain.
Single nuclei were extracted from snap-frozen mouse brain using the Nuclei Extraction Buffer in combination with the gentleMACS™ Octo Dissociator. Immediately after nuclei extraction, nuclei were stained using DRAQ5™ Staining Solution. The image shows an overlay of DRAQ5™ (pink) and brightfield.

Figure

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Single nuclei suspension from mouse kidney. Single nuclei were extracted from snap-frozen mouse kidney tissue using the Nuclei Extraction Buffer in combination with the gentleMACS Octo Dissociator. Immediately after nuclei extraction, nuclei were stained using DRAQ5™ Staining Solution. The image shows an overlay of DRAQ5™ (pink) and brightfield.
Single nuclei suspension from mouse kidney. Single nuclei were extracted from snap-frozen mouse kidney tissue using the Nuclei Extraction Buffer in combination with the gentleMACS Octo Dissociator. Immediately after nuclei extraction, nuclei were stained using DRAQ5™ Staining Solution. The image shows an overlay of DRAQ5™ (pink) and brightfield.

Specifications for Nuclei Extraction Buffer

Overview

The Nuclei Extraction Buffer is designed to gently, rapidly, and effectively generate single nuclei suspensions from fresh and frozen tissues. It has been developed for use with the gentleMACS Dissociators and C Tubes.

Detailed product information

Background information

The Nuclei Extraction Buffer allows for the preparation of single nuclei suspensions from a wide range of tissues types including brain, liver, heart, pancreas and kidney, as well as human and mouse solid tumors. Our method is optimized for the extraction of nuclei from snap-frozen tissue and it is also applicable for OCT embedded and fresh tissues.
The Nuclei Extraction Buffer is to be used with the gentleMACS Octo Dissociators and C Tubes and integrates tissue dissociation and cell lysis in a single step. You can extract nuclei in seven minutes at 4°C and run up to eight samples in parallel. Our standardized nuclei extraction method enables the generation of high quality nuclei suspensions to high yields.

Downstream applications

The nuclei extraction method is compatible with a wide variety of applications, including single nucleus gene expression analysis and single nucleus ATAC-seq. Some applications, e.g. single nucleus ATAC-seq, required nuclei permeabilization after extraction.
For details on the permeabilization, please follow the recommendation of the genomic application provider.

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