NuCLEAR™ Extraction Kit
SIGMA/NXTRACT - For mammalian tissue or cultured cells
Synonym: Nuclear Isolation Kit
Product Type: Chemical
Probe: Oct-1 Motif
Preparation Method: IGEPAL® Syringe
Figure 1. Gel Shift Assay of Oct-1.
Double stranded 32P-labeled Octamer motif oligonucleotide was incubated with either cytoplasmic (C) or nuclear extracts (N) of HeLa cells prepared using the CelLytic Nu-CLEAR Extraction kit. Two different methods, described in the protocol were utilized: Method 1 used NP-40 detergent; Method 2 relied on passages through a syringe instead of a detergent to extract the nuclei.
Preparation Method: IGEPAL® Syringe
Figure 1. Gel Shift Assay of Oct-1.
Double stranded 32P-labeled Octamer motif oligonucleotide was incubated with either cytoplasmic (C) or nuclear extracts (N) of HeLa cells prepared using the CelLytic Nu-CLEAR Extraction kit. Two different methods, described in the protocol were utilized: Method 1 used NP-40 detergent; Method 2 relied on passages through a syringe instead of a detergent to extract the nuclei.
Figure 2. Purity of Cytoplasmic and Nuclear Proteins.
The double stranded 32P-labeled Octamer motif oligonucleotide was incubated with either cytoplasmic fraction (C) or nuclear extract (N) prepared from HeLa, CHO, COS and PC-12 cells using the CelLytic-NuCLEAR extraction kit.
Probe: Octamer Motif
The double stranded 32P-labeled Octamer motif oligonucleotide was incubated with either cytoplasmic fraction (C) or nuclear extract (N) prepared from HeLa, CHO, COS and PC-12 cells using the CelLytic-NuCLEAR extraction kit.
Probe: Octamer Motif
| Quality Level | 300  |
| shipped in | dry ice |
| storage temp. | −20°C |
| technique(s) | protein extraction: suitable |
| western blot: suitable | |
| usage | kit sufficient for 10 extractions (1 ml packed cell volume) |
| kit sufficient for 100 extractions (100 μl packed cell volume) |
| Application: | NXTRACT kit was used to study the impact of salt on cardiac differential gene expression and coronary lesion in normotensive mineralocorticoid-treated mice. It was also used to test the therapeutic potential of andrographolide for treating endometriosis. |
| General description: | The procedure for the nuclear protein extraction method is to allow cells to swell with hypotonic buffer. The cells are then disrupted, the cytoplasmic fraction is removed, and the nuclear proteins are released from the nuclei by a high salt buffer. |
| Legal Information: | NuCLEAR is a trademark of Sigma-Aldrich Co. LLC |
| Other Notes: | A number of different procedures in the detailed technical bulletin enable the selection that best fits a particular application. For example, choose between detergent and non-detergent extraction of nuclear protein or between the standard hypotonic lysis buffer for most cell types and isotonic lysis buffer for fragile cells. In addition, the kit provides a procedure for salt reduction from the nuclear extract with dilution buffer. NuCLEAR offers the flexiblity you need for optimal protein extraction. Extracts can be prepared in less than 2 hours and are highly pure since there is little or no cross-contamination between nuclear and cytoplasmic extracts. |
| Other Notes: | Recommended Antibodies for Immunodetection L1293 , N2662 , AMAB90549 |
| Other Notes: | Within this kit is a complete system for preparing nuclear and cytoplasmic protein extracts from mammalian tissue or cultured cells. All reagents necessary for extraction are included. |
| Symbol |   GHS05,GHS09 |
| Signal word | Danger |
| Hazard statements | H290 - H314 - H410 |
| Precautionary statements | P273 - P280 - P303 + P361 + P353 - P304 + P340 + P310 - P305 + P351 + P338 + P310 - P391 |
| Hazard Codes | C |
| Risk Statements | 35 |
| Safety Statements | 26-36/37/39-45 |
| RIDADR | UN 3316 9 |
| Flash Point(F) | 188.6 °F - closed cup |
| Flash Point(C) | 87 °C - closed cup |
| Storage Temp. | −20°C |
| UNSPSC | 41105517 |
| Components | 3× Dilution and Equilibration Buffer 90 mL; Protease Inhibitor Cocktail 1 mL |