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Thermo Scientific™ Nitrocellulose Membranes, 0.2 μm
Description
Nitrocellulose is a popular binding matrix for western blotting because of its high affinity for proteins and compatibility with a variety of detection methods (western blotting, dot-blot assays, and other protein or nucleic acid methods). The 0.2-μm pore size is well-suited for western blotting, and this smaller pore size is ideal for the transfer of low molecular weight proteins or peptides and nucleic acids (<300 bp).
Features include:
100% pure nitrocellulose membrane
• Convenient—save time with pre-cut sheets
• Compatible with commonly used transfer conditions and detection methods such as staining, immunodetection, chemiluminescence, fluorescence, and radiolabeling
• High-binding affinity—provides excellent protein binding, blocks easily, and does not require pre-wetting with alcohol
• Low background—produce high-quality data with low background signal in both chemiluminescent and fluorescent western blotting
Specifications
Specifications
| Length (Metric) | 8 cm |
| Width (Metric) | 8 cm |
| Description | Nitrocellulose Membranes, 0.2 μm, 8 x 8 cm |
| Binding Capacity | ∼209 μg/cm2 of protein |
| Quantity | 15 Pre-cut Membranes |
| Format | Sheet |
| Material | Nitrocellulose |
| Pore Size | 0.2 μm |
| Content And Storage | Store membranes flat at ambient temperature, away from chemical vapors. Some solvent vapors may partially dissolve the membranes, which will disrupt the pore structure. Keep membranes out of direct sunlight. Sunlight may cause the nitrocellulose to dry and become brittle. Note: Nitrocellulose membrane filters are highly flammable. Keep away from heat and open flame. Flash point is approximately 2°C. Membranes can be sterilized by steam-autoclaving at 121°C. |
| Shipping Condition | Room Temperature |
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Frequently Asked Questions (FAQs)
For nitrocellulose or PVDF membrane following Western blot detection using a chemiluminescent or fluorescent substrate system: Following transfer, air dry the membrane and place in an envelope, preferably on top of a supported surface to keep the membrane flat. The blot can be stored indefinitely at -80 degrees C. When ready to reprobe, prewet the PVDF blot with alcohol for a few seconds, followed by a few rinses with pure water to reduce the alcohol concentration. Then proceed as normal with blocking step.
FOR STRIPPING/REPROBING OF MEMBRANES:
Harsh protocol (see NOTE below for modifications)
1) Submerge the membrane in stripping buffer (100 mM BME, 2% SDS, 62.5 mM Tris-HCl, pH 6.7) and incubate at 50 degrees C for 30 min with occasional agitation. If more stringent conditions necessary, incubate at 70 degrees C.
2) Wash 2 x 10 min in TBS-T/PBS-T at room temperature.
3) Block the membrane by immersing in 5% blocking reagent TBS-T or PBS-T for 1 hr at room temperature.
4) Immunodetection
NOTE: Often you don't need such harsh conditions to remove antibodies from their proteins. The stringency of one or several of the variables can be decreased: lower the temperature, decrease the time, less BME, less SDS, etc. An especially mild but still often effective stripping protocol is lower pH incubation. Example: pH 2.0 Tris 50-100 mM, 30-60 min incubation (you may do two incubations if you wish). Then rinse and block as usual. If you do not wish to re-use the membrane immediately after stripping, you can store the membrane in plastic wrap (wet, you do not want it to dry out). Another simple, mild stripping buffer is 0.1 M glycine•HCl (pH 2.5-3.0), incubation 30 min to 2 hrs room temperature or 37 degrees C, depending on the antibody.
Yes, both our PVDF and nitrocellulose membranes are compatible with the Li-COR instrument.
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