PerfectHyb™ Plus Hybridization Buffer
SIGMA/H7033 - for Northern and Southern blotting, solution
Synonym: hyb buffer; hyb solution; hybridization solution; northern hybridization buffer; southern hybridization buffer
Product Type: Chemical
Figure 1. Hybridization timecourse comparing PerfectHyb Plus with a conventional hybridization buffer (250 mM sodium phosphate buffer [pH 7.2], 7% SDS and 2mM EDTA). Northern blots containing ~2 μg human total RNA per lane were hybridized with ~200,000 cpm of 32P-labeled β-actin RNA probe in the indicated hybridization buffer. Blots were washed, imaged, and quantitated together on the Instant Imager.
Figure 2. Comparison of Northern blot hybridizations in different hybridization buffers for 1 and 20 hours. Northern blots containing ~2 μg human total RNA per lane were hybridized with ~200,000 cpm of 32P-labeled GAPDH RNA probe in the indicated hybridization buffer (Conventional buffer = 250 mM sodium phosphate buffer [pH 7.2], 7% SDS and 2mM EDTA). Blots were washed, imaged, and quantitated together on the Instant Imager.
Figure 3. Hybridization timecourse comparing PerfectHyb Plus with a conventional hybridization buffer (250 mM sodium phosphate buffer [pH 7.2], 7% SDS and 2 mM EDTA). Data obtained from blots shown in Figure 1 were plotted to show differences in the rates of hybridization in the two buffers. Signals equivalent to those obtained with overnight hybridizations in conventional buffers are obtained in ~3 hours using PerfectHyb Plus. In addition, PerfectHyb Plus achieves 80% of maximal signal within the first 6 hours of hybridization.
| form | solution |
| Quality Level | 200  |
| storage temp. | room temp |
| Application: | Suitable for use in hybridization buffers used in Northern blot, Southern blot, colony/plaque blots, and dot blots. |
| Features and Benefits: | • Fast and easy-to-use • Up to 20x more sensitive than conventional buffers • Doesn’t require blocking steps • Compatible with neutral and positively-charged membranes • Compatible with chemiluminescent, colorimetric , and DIG detection assays • Compatible with radioactive and non-radioactive probes • Compatible with single or double-stranded DNA, RNA, and oligonucleotide probes |
| General description: | Sigma’s PerfectHyb Plus is a ready-to-use hybridization buffer for Northern and Southern blot protocols. Use of PerfectHyb Plus can decrease hybridization time to as little as two hours. |
| Legal Information: | PerfectHyb is a trademark of Sigma-Aldrich Co. LLC |
| Other Notes: | A small amount of precipitate that has formed in the solution can be dissolved at 30-35°C with occasional gentle inversion. |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 2 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | room temp |
| UNSPSC | 12161700 |