Content Loading...
Content Loading...
Ordering Information
- : 2076
- : 1 kit
Product Images
BLOT-FastStain™ is a unique proprietary system for the reversible staining of proteins after transfer to nitrocellulose or PVDF membranes. The stainin...
BLOT - FastStain
Description:
BLOT - FastStain
Trade Name:
Chemicon (Millipore)
Qty/Pk:
1 kit
Product Overview:
The Millipore BLOT-FastStain™ is a unique proprietary system for the reversible staining of proteins after transfer to nitrocellulose or PVDF membranes. The staining procedure takes 10 minutes. BLOT-FastStain™ detects only proteins (BLOT-FastStain™ does not stain nucleic acids) and leaves the background absolutely untouched and brilliant white leading to exceptional band visibility. The lower detection limit of BLOT-FastStain™ is 0.5 ng/band (BSA). The staining color is indigo blue. BLOT-FastStain™ can be removed from transfer membranes in 10 minutes without affecting their biological or immunological properties. After removing the stain, Western blotting can proceed with no changes in protocol. BLOT-FastStain™ therefore allows you to measure the transfer of proteins before potentially wasting expensive and sometimes irreplaceable antibodies. Other applications include detecting size fractionated proteins for later antibody generation or sequencing.
Key Applications:
Western Blotting
Application Notes:
STAINING PROTOCOL:
These instructions are for 50 mL (enough for a single 12 x 12 cm2 membrane). Increase volumes with larger gels; it is not recommended to use volumes of less than 50 mL.
1. Dilute Reagent A: Fixer 10 fold by adding 5 mL of the fixer to 45 mL of deionized water. Place transfer membrane (nitrocellulose or PVDF) into a tray containing the diluted fixer. Incubate for 2-3 minutes with gentle shaking.
2. Dilute Reagent B: Developer 4 fold by adding 10 mL of the Developer to 30 mL deionized water. Transfer the membrane (nitrocellulose or PVDF) into the tray containing the diluted Developer. Incubate for 1 minute with gentle shaking.
3. Transfer the Developer tray into a refrigerator for protein bands to develop. Shaking at this stage is not recommended.
4. Incubate until protein bands can be detected and reach their desired intensity. The best results will be achieved if the membranes are developed in the cold, dark, and without shaking or disturbing them.
5. Protein bands will be visible within 10 minutes of incubation and reaches full intensity within 30 minutes. Developing time depends on the concentration of protein on the surface of the membrane. After staining, membrane may be left in developer for several days.
Background Staining: Background staining depends on the types of membrane used for protein transfer. Nitrocellulose gives the most clear and brilliant white background. Some brands of PVDF membranes may give higher background. Background staining can be removed by shaking the membrane 5-10 minutes in cold water (too much washing may de-stain the protein bands).
DESTAINING:
To de-stain, simply place the membrane in warm deionized water (40-50°C) and shake in bright light for 10 minutes or until stain disappears. Wash twice, 10 minutes each, with deionized water.
These instructions are for 50 mL (enough for a single 12 x 12 cm2 membrane). Increase volumes with larger gels; it is not recommended to use volumes of less than 50 mL.
1. Dilute Reagent A: Fixer 10 fold by adding 5 mL of the fixer to 45 mL of deionized water. Place transfer membrane (nitrocellulose or PVDF) into a tray containing the diluted fixer. Incubate for 2-3 minutes with gentle shaking.
2. Dilute Reagent B: Developer 4 fold by adding 10 mL of the Developer to 30 mL deionized water. Transfer the membrane (nitrocellulose or PVDF) into the tray containing the diluted Developer. Incubate for 1 minute with gentle shaking.
3. Transfer the Developer tray into a refrigerator for protein bands to develop. Shaking at this stage is not recommended.
4. Incubate until protein bands can be detected and reach their desired intensity. The best results will be achieved if the membranes are developed in the cold, dark, and without shaking or disturbing them.
5. Protein bands will be visible within 10 minutes of incubation and reaches full intensity within 30 minutes. Developing time depends on the concentration of protein on the surface of the membrane. After staining, membrane may be left in developer for several days.
Background Staining: Background staining depends on the types of membrane used for protein transfer. Nitrocellulose gives the most clear and brilliant white background. Some brands of PVDF membranes may give higher background. Background staining can be removed by shaking the membrane 5-10 minutes in cold water (too much washing may de-stain the protein bands).
DESTAINING:
To de-stain, simply place the membrane in warm deionized water (40-50°C) and shake in bright light for 10 minutes or until stain disappears. Wash twice, 10 minutes each, with deionized water.
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Components:
- Reagent A: Fixer, 120 mL
- Reagent B: Developer, 2 X 125 mL
Brand Family:
Chemicon
Storage Conditions:
Store kit components at room temperature for up to twelve months from date of purchase. To avoid possible microbial contamination, dispense all solutions aseptically. Reagents must not be stored in cold temperatures.
Warning: Some reagents are weakly acidic. However, no components are hazardous; no extraordinary precautions other than general laboratory procedures are necessary (e.g., dispense aseptically, avoid any contact with eyes or skin).
Warning: Some reagents are weakly acidic. However, no components are hazardous; no extraordinary precautions other than general laboratory procedures are necessary (e.g., dispense aseptically, avoid any contact with eyes or skin).
Reagent Type:
Western Blotting Reagents
Product Name:
BLOT - FastStain