Material needed but not supplied
1) Microwave oven.
2) Microwavable plastic box.
Features and Applications
Sensitivity: Detection of protein as low as 25 ng (BSA) can be achieved in 10 min.Sensitivity can be extended to 10 ng for some proteins (such as BSA) with additional staining treatment and washing steps.
Applications:Detection of proteins in native and SDS-PAGE gels.
Room temperature in a lightproof bottle.
Warning: The instructions described below involve handing of “hot” liquid. Follow the directions with caution to prevent burn.
1) Place a standard mini gel in a microwavable plastic box with 250 ml of deionized water (or sufficient amount) to cover the gel. Loosely cover the box and microwave the gel for 3 min. (high setting).
2) Discard the water. Repeat step 1.
3) Optional step: For maximum detection sensitivity, repeat step 1 two more times (to a total of 4 water washings).
4) Submerge the gel in 50 ml (or sufficient amount) of BlueFast solution. Loosely cover the box and microwave the gel for 2 min. (high setting). For maximum detection, submerge the gel with 100 ml of BlueFast solution, and microwave the gel for 4 min.
5) Discard stain. Add 250 ml of deionized water. Loosely cover the box and microwave the gel for 2 min. (high setting). Discard wash. Add 250 ml of deionized water.
6) Optional step: For maximum detection sensitivity, repeat step 5.
7) Optional step :BlueFast leaves a light-blue background, which does not affect the sensitivity of detection on the gel. To remove it completely, wash the gel 3 times, 30 min. each with rocking, with 250 ml of deionized water, or simply leave the gel in water overnight.
Q: What is BlueFast?
A: BlueFast is a proprietary formulation of coomassie blue G-250 stain solution.Its unique formula works its magic with heat to speed up the process of protein staining with high detection sensitivity.
Q: What is the sensitivity of BlueFast?
A: BlueFast works best for detection of proteins 15 k Da and above.It’s detection sensitivity varies from protein to protein depending on their composition.In general,BlueFast detects as low as 25 ng of BSA in native and SDS-PAGE within 10 minutes using the standard procedure.It is possible to increase the sensitivity of BlueFast detection to as low as 10 ng of BSA.To achieve this, repeat the water treatment step two more times (to a total of 4 treatments), increase the BlueFast staining step from 2 to 4 minutes, and repeat the post staining wash with heating one more time.
Q: How does the detection sensitivity of BlueFast compared to traditional coomassie blue staining method and silver staining method?
A: BlueFast is based on colloidal coomassie blue staining technology so it is more sensitive than the traditional coomassie blue staining method.Silver staining provides higher detection sensitivity for some proteins than BlueFast.However, silver staining is more difficult to work with and its high detection sensitivity is not universally applicable to all proteins.
Q: How does the water treatment work?
A: Boiling of the gel in water prior to the BlueFast step removes substances that can inhibit the interaction between BlueFastä and proteins in the gel.
Q: What is the effect of boiling on polyacrylamide gel?
A: Boiling does not destroy the polyacrylamide gel.The only beneficial effect of boiling on the polyacrylamide gel is it fixes the gel so it will not expand in water.
Q: Why proteins detected by BlueFast show a lighter shade of blue compared to the color seen when using the traditional coomassie blue staining method?
A: The unique formulation of BlueFastä yields a lighter blue-colored stained protein than that produced by the traditional coomassie blue staining method.This does not decrease the sensitivity of protein detection.
Q: Can BlueFast be reused?
A: Yes, but we don’t recommend it unless one is working with abundant amount of proteins.Detection sensitivity decreases 50% when BlueFast is used the second time.Therefore, unless there is a high concentration of protein in the gel, we do not recommend using BlueFast a second time.
Q: Can proteins be transferred to a membrane from a gel after they have been stained by BlueFast?
A: No for most proteins.Once proteins are stained by BlueFast, they can not be transferred.It is misleading to see blue bands on a membrane if one attempts to transfer BlueFast stained proteins from gel.The transferred blue bands are coomassie blue dyes.Since protein detection by BlueFast is so rapid, we recommend running two identical gels simultaneously. Leave one gel in the electrophoresis apparatus, stain the second gel with BlueFast and obtain the results within 10 minutes.Based on the results of the stained gel, the user then can decide whether to transfer the unstained gel to a membrane.
Q: Can I remove the light-blue background produced by BlueFast completely?
A: BlueFast leaves a light-blue background, which does not affect the sensitivity of detection on the gel.To completely remove the light-blue background, one can repeat the post staining wash step with heating several times, leave the gel in 500 ml of water and change the water hourly, or simply leave the gel in water overnight.
Q: Can I treat the BlueFast stained gel with gel drying reagents containing glycerol and ethanol?
NOTE: We recommend washing the gels twice (10 minutes each) with water and then processing the gel with gel drying reagents.
Q: Can I sequence proteins stained by BlueFast using mass spec?
A: Yes. Excise the protein of interested from the stained gel.Wash the excised gel 3 times with water (one hour each) or leave it in water overnight.Then proceed to the sequencing procedure.
MATERIAL SAFETY DATA SHEET
TRADE NAME: BlueFast Protein Staining Solution
DATE OF ISSUE: February 8,2017
SKU#: 1900011002 (100 ml), 1900010013 (1000 ml)
TRADE NAME:BlueFast Protein Staining Solution
Manufacturer: Eton Bioscience Inc., 5820 Oberlin Drive, Suite 108, San Diego, CA 92121
Emergency Telephone: 800.758.1630
SECTION II: COMPOSITION / INFORMATION ON INGREDIENTS
Brilliant Blue G250
HAZARDOUS COMPONENTS: The product contains no substances which at their given concentration, are considered to be hazardous to health.
SECTION III:HAZARD IDENTIFICATION
Irritating to eyes and skin. May cause sensitization by skin contact.
SECTION IV:FIRST AID MEASURES
General Information: Remove and wash contaminated clothing promptly.
IN CASE OF INHALATION: Supply fresh air and then consult a doctor.
IN CASE OF EYE CONTACT: Immediately flush eyes with copious amounts of water for 15 minutes.
IN CASE OF SKIN CONTACT: Immediately wash skin with soap and copious amounts of water and consult a doctor. Remove contaminated clothing and shoes. Wash clothing before reuse. Clean shoes thoroughly before reuse.
IF SWALLOWED: Rinse mouth with copious amounts of water. Do not induce vomiting. Never give anything by mouth to a unconscious person.Then consult a doctor.
SECTION V:FIREFIGHTING MEASURES
Suitable: Use Dry chemical, water spray, carbon dioxide or foam.
SECTION VI:ACCIDENTAL RELEASE MEASURES
PRECAUTIONARY MEASURES:No special measures required.
CLEAN-UP PROCEDURES:No special measures required.
SECTION VII:HANDLING AND STORAGE
HANDLING: Avoid inhalation of vapour or mist. Keep away fro sources of ignition. No smoking.
STORAGE: Keep container tightly closed in a dry and well-ventilated place.
SECTION VIII:EXPOSURE CONTROLS / PERSONAL PROTECTION
The routine precautionary measures for handling chemicals should be followed such as wearing chemical safety goggles, rubber gloves, rubber boots, NIOSH / MSHA-Approved respirator when handling chemicals and wash thoroughly after handling.Handle chemicals with care and avoid contact with eyes, skin, or on clothing.Use safety shower and eye bath as first aid measures if exposed to chemicals.
SECTION IX:PHYSICAL AND CHEMICAL PROPERTIES
PHYSICAL STATE / FORM: Liquid
TRADE NAME: Bluefast Protein Staining Solution
pH FACTOR: (20°C) 2.5-3.5
VISCOSITY: (20°C) N/A
MELTING POINT: N/A
BOILING POINT: N/A
IGNITION TEMPERATURE: N/A
EXPLOSION LEVEL: LOWER: N/A
VAPOR PRESSURE: (20°C) N/A
SPECIFIC GRAVITY: (20°C) N/A
SOLUBILITY IN WATER: (20°C) Soluble
SECTION X:STABILITY AND REACTIVITY
SUBSTANCES TO BE AVOIDED: Oxidizing agents.
DANGEROUS REACTIONS: No dangerous reactions known.
HAZARDOUS, COMBUSTION, OR DECOMPOSITION PRODUCTS:
Under normal conditions of storage and handling, hazardous, combustion or decomposition products should not be produced.
SECTION XI:TOXICOLOGICAL INFORMATION
TOXICITY DATA: N/A
ACUTE EFFECTS INHALATION: N/A.
EYE CONTACT: May cause eye irritation.
SKIN CONTACT: May cause skin irritation.
INGESTION: May be harmful if swallowed.
PROLONGED EXPOSURE: N/A
CHRONIC EFFECTS: N/A
RTECS number: N/A
The product should be handled with routine precautionary measures for handling chemicals. Additional harmful properties cannot be ruled out.
SECTION XII:ECOLOGICAL INFORMATION
No information available.
SECTION XIII:DISPOSAL CONSIDERATIONS
Chemical waste generators must determine whether a discarded chemical is classified as a hazardous waste. US EPA guidelines for the classification determination are listed in 40 CFR Parts 261.3. Additionally, waste generators must consult state and local hazardous waste regulations to ensure complete and accurate classification.
SECTION XIV:TRANSPORT INFORMATION
UN Number:1170 Class:3 Packing group: III
Proper Shipping Name: Ethanol Solutions
SECTION XV:REGULATORY INFORMATION
No information available.
SECTION XVI:OTHER INFORMATION
The information above is believed to be accurate and represents the best information currently available to us. However, we make no warranty of merchantability or any other warranty, express or implied, with respect to such information, and we assume no liability resulting from its use. Users should make their own investigations to determine the suitability of the information for their particular purposes. Eton Bioscience Inc. shall not be held liable for any damages or other consequences resulting from handling or from contact with the above product.