CUTANA™ Nuclei Extraction Buffer
26 in stock
Powered by BiozCUTANA™ Nuclei Extraction Buffer is the essential reagent for harvesting nuclei from cultured cells and tissues for use in CUT&RUN and CUT&Tag assays. This buffer is expected to be broadly compatible with eukaryotic cells and tissues. Below are examples of various cells and tissue types that have undergone successful nuclei isolation using this buffer:
| mouse NIH3T3 fibroblast cells | human A549 non-small cell lung cancer (NSCLC) cells |
| human K562 leukemia cells | human NCI-H1299 non-small cell lung cancer (NSCLS cells) |
| human bone marrow derived macrophages | human TIG-1 fetal lung cells |
| human monocyte derived macrophages | human LoVo colorectal cancer cells |
| human MV-4-11 macrophage cells | human LNCaP prostate carcinoma cells |
| human SUM149 triple negative breast cancer (TNBC) cells | human renal primary cells |
| human GM24385 B-lymphocyte (aka HG002) cells | human peripheral blood mononuclear cells (PBMCs) |
| human MCF7 breast cancer cells | human intestinal tissue |
| human MDA-MB-231 breast cancer cells | human HEPM embryonic cells |
| human SK-MEL-2 melanoma cells |
The reagent is prepared by supplementing Pre-Nuclei Extraction Buffer with protease inhibitor and spermidine fresh on the day of use. Utilize this buffer in EpiCypher’s CUTANA™ Nuclei Extraction Protocol for CUT&RUN and CUT&Tag to obtain the highest quality nuclei for your genomic mapping assay.
Figure 1: Nuclei Extraction
Store the Pre-Nuclei Extraction Buffer at 4ºC and the 1 M Spermidine at -20ºC. Components are stable for 6 months upon date of receipt.
Prepare Nuclei Extraction Buffer FRESH on day of use as outlined below. This master mix includes 20% extra volume to account for pipetting error; no additional buffer volume is needed.
Per CUT&RUN or CUT&Tag reaction, combine:
235 µL Pre-Nuclei Extraction Buffer (21-1026a)
0.13 µL 1M Spermidine (21-1026b)*
9.8 µL 25X Protease Inhibitor (Prepare using CUTANA™ Protease Inhibitor Tablets, 21-1027)**
*If preparing buffer for fewer than 8x reactions, dilute the 1M Spermidine stock 1:10 in molecular biology-grade water and add 1.3 µL per reaction.
**Leftover 25X Protease Inhibitor can be stored for 12 weeks at -20ºC
Follow instructions detailed in EpiCypher’s CUTANA™ Nuclei Extraction Protocol for CUT&RUN and CUT&Tag to isolate nuclei from suspension and adherent cells. The protocol includes key quality control checks to ensure nuclei are high quality before starting CUT&RUN or CUT&Tag.
| Item | Cat. No. |
|---|---|
| CUTANA™ Pre-Nuclei Extraction Buffer | 21-1026a |
| 1 M Spermidine | 21-1026b |
Required Materials Not Supplied
Protease inhibitor is not included. EpiCypher suggests using CUTANA™ Protease Inhibitor Tablets (21-1027).
| Item | Cat. No. |
|---|---|
| CUTANA™ ChIC/CUT&RUN Kit | 14-1048 |
| CUTANA™ pAG-MNase for ChIC/CUT&RUN | 15-1016/15-1116 |
| CUTANA™ pAG-Tn5 for CUT&Tag | 15-1017/15-1117 |
| CUTANA™ Protease Inhibitor Tablets | 21-1027 |
NOTE:
CUTANA™ CUT&Tag Kit (14-1102/14-1103) already contains Nuclei Extraction Buffer