Nitrocellulose Membrane (NC) - BioReagent, for Northern and Southern blotting, for western blot, Aperture: 0.45 (μm), size: 100x100mm

Cat. No.: N766351

GRADE & PURITY BioReagent, for Northern and Southern blotting, for Western blot, Aperture: 0.45 (μm), size: 100x100mm

Synonyms

Nitrocellulose transfer membrane

Storage

Room temperature

Shipped In

Normal

Application

Northern blotting, Southern blotting, WB

Descripción general

PVDF transfer membrane is a microporous polyvinylidene fluoride (PVDF) membrane used for transferring proteins from various gel matrices. It offers better handling properties compared to nitrocellulose membranes. This hydrophobic membrane is available in two pore sizes, 0.45 μm and 0.22 μm, and can bind proteins with a wide range of molecular weights.

NC transfer membrane is a microporous nitrocellulose membrane designed for protein transfer from diverse gel matrices. Unlike PVDF membranes, NC membranes are hydrophilic and do not require pre-wetting activation. Supplied in 0.45 μm and 0.22 μm pore sizes, this hydrophilic membrane is capable of binding proteins spanning a broad molecular weight range.

The 0.45 μm pore size is suitable for most proteins and delivers high detection sensitivity. The 0.22 μm pore size is optimized for proteins with molecular weights below 20 kDa, enabling superior adsorption and retention of small-molecular-weight proteins. Aladdin PVDF and NC transfer membranes feature excellent protein retention, robust physical strength, and broad chemical compatibility, making them well-suited for immunoassays.

Comparison of performance and applications for each catalog number of Aladdin PVDF and NC transfer membranes:

Precautions:
1. Always wear gloves when handling the membrane to avoid leaving fingerprints.
2. Use blunt forceps to prevent membrane damage.
3. Keep the protective light blue backing paper attached to the membrane during cutting or handling; discard the backing paper prior to membrane wetting.
4. Handle the membrane carefully to avoid scratches on its surface and do not fold it.
5. Hydrophobic PVDF transfer membranes must be pre-wetted with methanol solution, while hydrophilic NC transfer membranes require no pre-wetting. The membrane will turn translucent from opaque once wetted.
6. After protein transfer, rinse the blot with ultrapure water to remove residual contaminants.
7. Blotted membranes can be air-dried and stored at 4°C for several months for subsequent use, or used immediately.
Instructions for Use:
I. Membrane Wetting
1. Immerse the dry membrane in methanol for 10–20 seconds, or until it turns uniformly translucent gray from opaque white.
2. Submerge the membrane in ultrapure water for 1–2 minutes to displace methanol. (These two steps are not required for NC membranes.)
3. Equilibrate the membrane in transfer buffer for 2–3 minutes or until ready for use.
Note: Do not allow the membrane to dry out once wetted. It may remain submerged in buffer until protein transfer. If the membrane dries out (turns opaque white), even partially, repeat the wetting procedure (Steps 1–3).
II. Semi-Dry Transfer
1. Separate protein samples on a polyacrylamide gel.
2. Immerse the gel in transfer buffer and equilibrate for 10–15 minutes.
3. Assemble the transfer stack following the manufacturer’s instructions for your transfer unit.
Note: To ensure uniform protein transfer, gently roll a clean pipette or blot roller over the surface of each layer in the stack. Do not apply excessive pressure, as this may damage the gel and membrane.
4. Perform protein transfer in accordance with the transfer equipment manufacturer’s guidelines.
5. Remove the blot from the transfer system and briefly rinse the membrane with ultrapure water to eliminate gel debris. The blot can be air-dried for storage or used immediately for immunodetection.
Note: Drying the blot prior to immunodetection may improve binding of certain proteins and reduce background signal.
III. Standard Immunodetection
1. If the blot has dried, rewet it in methanol for 15 seconds or until it turns translucent gray from opaque white.
2. Rinse the blot with ultrapure water for 1 minute.
3. Place the blot in blocking buffer and incubate with gentle agitation for 1 hour. Dilute the primary antibody using commercial antibody diluent, wash buffer or blocking buffer.
4. Submerge the blot in diluted primary antibody solution and incubate at room temperature for 1 hour with gentle agitation (or incubate overnight at 4°C).
5. Wash the blot 3–5 times with wash buffer (buffer supplemented with Tween-20 surfactant, i.e., TBST or PBST), 5 minutes per wash. Prepare the diluted secondary antibody with commercial antibody diluent, wash buffer or blocking buffer.
6. Immerse the blot in diluted enzyme-conjugated secondary antibody solution and incubate at room temperature for 1 hour.
7. Wash the blot 3–5 times with wash buffer, 5 minutes each time.
8. Transfer the blotting membrane to a clean container and add the appropriate detection reagent.
9. Incubate for 1–5 minutes following the detection reagent manufacturer’s instructions.
10. For HRP or AP chemiluminescence reagents, expose the blot to X-ray film or capture images with a digital imaging system. For colorimetric detection, add the reagent and wait for signals to develop.

Specifications

Sinónimos

Nitrocellulose transfer membrane

Especificaciones y pureza

BioReagent, for Northern and Southern blotting, for western blot, Aperture: 0.45 (μm), size: 100x100mm

Estabilidad y almacenamiento

Store at room temperature for 24 months.

Condiciones de almacenamiento de almacenamiento

Room temperature

Enviado en

Normal

Grado

BioReagent, for Northern and Southern blotting, for Western blot

Imágenes

Western Blot: Nitrocellulose Membrane (NC) (N766351)

Nitrocellulose Membrane (NC) (N766351) - Western Blot 

The Western Blot performance of this nitrocellulose membrane (NC membrane) is demonstrated in the image. Cell lysates (MCF7, NIH/3T3, etc.) were separated by SDS‑PAGE, transferred to the NC membrane, and visualized after antibody incubation. Specific and distinct bands are observed across a broad molecular weight range of 10 kDa to 250 kDa (marker shown). With high protein‑binding capacity, ultra‑low background, sharp band resolution, and excellent signal‑to‑noise ratio, this membrane reliably supports detection of proteins across a wide molecular weight spectrum.

Propiedades químicas y físicas

• Sensibilidad: Light & Heat sensitive