| Field | Specification |
|---|---|
| Mfr No | |
| Detection Method | |
| Product Type |
Product Overview
The U-Blot® NC Transfer Membrane is a nitrocellulose microporous membrane designed for protein transfer from various gel matrices. Compared to PVDF membranes, NC membranes are hydrophilic and require no prior hydrophilic treatment. It is available in two pore-size variants (0.45 µm and 0.2 µm), enabling binding of proteins across a wide molecular-weight range.
The 0.45 µm pore size is suitable for most proteins and offers high detection sensitivity, while the 0.2 µm pore size is ideal for proteins with a molecular weight below 20,000, enabling superior adsorption and retention of small-molecule proteins. The membrane exhibits excellent protein retention, high physical strength, and broad chemical compatibility, making it well-suited for immunological assays.
Specifications
| Material | Nitrocellulose |
|---|---|
| Membrane Property | Hydrophilic |
| Pore Sizes | 0.2 µm and 0.45 µm |
| Protein Binding Capacity (BSA) | >160 µg/cm² (0.2 µm); >140 µg/cm² (0.45 µm) |
| Format / Dimensions | 30 cm x 3 m |
| Detection Methods | Chemiluminescent, Colorimetric, Radioactive |
| Applications | Western Blot, Northern Blot, Southern Blot |
Applications
Suitable for Western blot, Northern blot, and Southern blot workflows, with compatibility across chemiluminescent, colorimetric, and radioactive detection systems.
Compatibility & Usage Notes
Select the 0.45 µm pore size for most proteins and high sensitivity; choose the 0.2 µm pore size for proteins below ~20 kDa where superior retention of small proteins is needed.
Safety & Handling
Transfer Protocol
- Separate protein samples and molecular-weight markers by SDS-PAGE.
- Cut and equilibrate the gel in cold transfer buffer for ~20 minutes; cut membrane and filter paper to size.
- Pre-wet the NC membrane in ultrapure water for ~20 seconds, then equilibrate in cold transfer buffer for 2-3 minutes.
- Assemble the transfer sandwich: clamp -> sponge pad -> filter paper -> pre-wetted membrane -> pre-wetted gel -> filter paper -> sponge pad -> clamp, avoiding wrinkles and bubbles.
- Set the operating conditions per your transfer device and start the transfer.
- After transfer, remove the blot, rinse briefly with ultrapure water, then air-dry for storage or proceed directly to immunodetection. Note: drying before immunodetection may improve binding of some proteins and reduce background.
Handling Instructions
- Wear gloves to prevent fingerprint contamination.
- Use blunt forceps to prevent membrane damage.
- Keep the light-blue protective paper with the membrane while cutting; discard it after wetting.
- Handle carefully to avoid scratching the surface and do not fold the membrane.
- After transfer, rinse the blot with ultrapure water to remove residual impurities.
- The blot can be air-dried and stored at 4°C for several months, or used immediately.
Recommended Membrane by Application
- General immunological testing → PVDF or NC Transfer Membrane (0.45 µm)
- Amino-acid analysis → PVDF Transfer Membrane (0.45 µm)
- Detection of low-abundance or small-molecular-weight proteins → PVDF or NC Transfer Membrane (0.2 µm)
- Sequencing of low-abundance or low-MW proteins → PVDF Transfer Membrane (0.2 µm)
The 0.45 µm pore size suits most proteins and offers high detection sensitivity, while the 0.2 µm pore size is ideal for proteins below about 20 kDa, giving superior retention of small proteins.
It is hydrophilic and requires no methanol pre-treatment; pre-wet briefly in ultrapure water, then equilibrate in cold transfer buffer.
The membrane is compatible with chemiluminescent, colorimetric, and radioactive detection systems.
Always wear gloves and use blunt forceps, avoid folding or scratching the surface, and keep the membrane wet throughout immunodetection.
The membrane is supplied in roll format with dimensions of 30 cm x 3 m, in 0.2 µm and 0.45 µm pore-size variants.
BioHippo can source custom configurations, bulk volumes, and special packaging for this product through our manufacturing partner. Contact us with your requirements for a quote and lead time.
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