UltraBio™ SDS-PAGE Protein Staining and Loading Buffer

Cat. No.: S751602

GRADE & PURITY 5X, Odorless

Overview

Aladdin's UltraBio™ SDS-PAGE Protein Staining and Loading Buffer (5X, Odorless) is used for loading protein samples onto SDS-PAGE gels for electrophoresis. It contains an odorless reducing reagent and a safe and nontoxic fluorescent dye that stains protein in purple during the boiling process and emits orange fluorescence under a UV or blue light. Post-run staining and destaining of SDS-GAGE gels are eliminated.This product is non-hazardous and convenient to use. Add 1/4 volume of this product to your protein sample, mix well and boil at 95-100℃ for 5 minutes. Proteins are then stained purple and emit orange fluorescence. All procedures can be performed under normal lighting without additional protection from light.The small and uncharged fluorescent molecules contained in UltraBio™ SDS-PAGE Protein Staining and Loading Buffer (5X, Odorless) do not affect the migration rate of proteins and the resulting electrophoresis pattern.Electrophoresis results can be visualized immediately after the run. Proteins of more than 500ng can be directly observed by naked eyes. The fluorescence of proteins in small quantities can be observed in a dark room using UV or blue light.This product is compatible with western blotting applications. Proteins in the SDS-PAGE gel can be transferred to PVDF or nitrocellulose membranes after electrophoresis. The purple band or orange fluorescence of proteins could also be observed on the membrane after transfer. The sensitivity of Western Blot detection is not affected by this product, which has been confirmed through testing a variety of commonly used antibodies. The possibility of affecting the detection of particular proteins can not be excluded.The background of gel after electrophoresis is low and free fluorescent dye does not affect observation and detection of proteins. The free fluorescent dye and the bromophenol blue tracking dye comigrate together to the end of the gel to ensure a clean background of the gel.This product has high sensitivity and can detect protein down to 25ng. A single protein band with more than 500ng can be observed obviously by naked eyes (Figure 1), and a 200ng protein band is faintly visible. Under a UV or blue light, proteins higher than 25ng can be observed (Figure 2).This product is compatible with the commonly used Tris-Glycine and Hepes gel buffer systems.The fluorescent signal is stable. The fluorescence of protein on SDS-PAGE gels is stable at 4ºC for at least 5 hours. If transferred immediately after electrophoresis, the fluorescence of protein on membranes is stable at 4ºC for at least 24 hours.This product is more secure to use. An odorless, more stable water-soluble reducing reagent with a similar reducing capability to dithiothreitol (DTT) or 2-mercaptoethanol is used in this product, making the product odorless during normal use or heating.Main Applications

Application：

Rough comparison of protein quantity in each lane, especially for detection of tissue samples using Western Blot; Quick and convenient examination of protein induction, expression, and purification.Figure 1. Staining result of different amounts of BSA run with ’s InstantView™ Protein Staining and Loading Buffer (5X, Odorless) . Mix different amounts of BSA proteins, as indicated in the figure, with the appropriate amount of this loading buffer, boil at 100ºC for 5 min, followed by electrophoresis. This photo was taken immediately after electrophoresis. The 500ng band can be observed obviously by naked eyes and the 200ng protein band is faintly visible.Figure 2. Images of protein samples run with the InstantView™ Protein Staining and Loading Buffer (5X, Odorless) on SDS-PAGE gels. A and B. Mix different amounts of BSA proteins, as indicated in the figure, with the appropriate amount of this loading buffer, boil at 100ºC for 5 min, followed by electrophoresis. Picture A was taken immediately after electrophoresis under a UV light and Picture B was taken immediately after electrophoresis under a blue light. C. Stain result of bacteria cell lysate induced by IPTG to express a 35 kD target protein. Fluorescence was observed under UV light (left) after electrophoresis or under blue light (middle). The staining result with Coomassie Brilliant Blue is shown on the right.Packing List：ItemComponentQuantityP0281S-1InstantView™ Protein Staining DyeFor 1mlP0281S-2InstantView™ Protein Loading Buffer (5X, Odorless)1mlManual-1 copyStorage Conditions：Store at -20℃ for up to 1 year, 4℃ for up to 3 months, or room temperature for up to 2 weeks. The InstantView™ Protein Staining Dye is light-sensitive and should be protected from light.Precautions：Protein samples after mixing with the InstantView™ SDS-PAGE Protein Staining and Loading Buffer (5X, Odorless) should be boiled at 95℃ for 5 minutes. Too high temperature (e.g., 100℃) or too long time of boiling (e.g., more than 15 minutes) could cause protein degradation or aberrance of the tracking dye.InstantView™ SDS-PAGE Protein Staining and Loading Buffer (5X, Odorless) contains an odorless reducing reagent, but with a similar reducing capability to toxic 2-mercaptoethanol and irritating DTT.The prepared InstantView™ SDS-PAGE Protein Staining and Loading Buffer (5X, Odorless) must be melted completely prior to use.Lower pH significantly affects protein staining by the InstantView™ SDS-PAGE Protein Staining and Loading Buffer. When the pH of protein samples is below 5, we recommend adjusting the pH to above 7.Most prestained and non-prestained protein molecular weight standards are incompatible with this product. Non-prestained protein molecular weight standards without any loading buffers or dissolved in water or saline can be mixed with an appropriate amount of InstantView™ SDS-PAGE Protein Staining and Loading Buffer (5X, Odorless) to produce protein molecular weight standards that emit orange fluorescence.Proteins stained by this product are not recommended for subsequent protein sequencing, protein profiling, or antibody preparation.InstantView™ SDS-PAGE Protein Staining and Loading Buffer (5X, Odorless) should be kept at -20ºC in the dark. Its staining performance reduces after 2-3 weeks at room temperature because of oxidation, which can be avoided by adding DTT in the loading buffer at a concentration of no more than 20mM.This product is hazardous. Please handle with care to avoid inhalation or direct contact with the human body.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation.

Precautions：

Protein samples after mixing with the InstantView™ SDS-PAGE Protein Staining and Loading Buffer (5X, Odorless) should be boiled at 95℃ for 5 minutes. Too high temperature (e.g., 100℃) or too long time of boiling (e.g., more than 15 minutes) could cause protein degradation or aberrance of the tracking dye.InstantView™ SDS-PAGE Protein Staining and Loading Buffer (5X, Odorless) contains an odorless reducing reagent, but with a similar reducing capability to toxic 2-mercaptoethanol and irritating DTT.The prepared InstantView™ SDS-PAGE Protein Staining and Loading Buffer (5X, Odorless) must be melted completely prior to use.Lower pH significantly affects protein staining by the InstantView™ SDS-PAGE Protein Staining and Loading Buffer. When the pH of protein samples is below 5, we recommend adjusting the pH to above 7.Most prestained and non-prestained protein molecular weight standards are incompatible with this product. Non-prestained protein molecular weight standards without any loading buffers or dissolved in water or saline can be mixed with an appropriate amount of InstantView™ SDS-PAGE Protein Staining and Loading Buffer (5X, Odorless) to produce protein molecular weight standards that emit orange fluorescence.Proteins stained by this product are not recommended for subsequent protein sequencing, protein profiling, or antibody preparation.InstantView™ SDS-PAGE Protein Staining and Loading Buffer (5X, Odorless) should be kept at -20ºC in the dark. Its staining performance reduces after 2-3 weeks at room temperature because of oxidation, which can be avoided by adding DTT in the loading buffer at a concentration of no more than 20mM.This product is hazardous. Please handle with care to avoid inhalation or direct contact with the human body.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation.

Instructions for Use：

1. Prepare the InstantView™ SDS-PAGE Protein Staining and Loading Buffer (5X, Odorless).a. Centrifuge the InstantView™ Protein Staining Dye to the bottom of the tube at 5,000-10,000g for 10-20 seconds.b. Thaw the InstantView™ Protein Loading Buffer (5X, Odorless) at room temperature. SDS contained in the loading buffer precipitates when stored cold, but will be resolubilized at room temperature.c. Add 1ml of InstantView™ Protein Loading Buffer (5X, Odorless) into the InstantView™ Protein Staining Dye, mix and dissolve the dye thoroughly to generate the InstantView™ SDS-PAGE Protein Staining and Loading Buffer (5X, Odorless).Note: The prepared InstantView™ SDS-PAGE Protein Staining and Loading Buffer (5X, Odorless) should be used immediately or can be kept at -20ºC in the dark for up to 2 months.2. Mix the Protein Staining and Loading Buffer (5X, Odorless) and protein samples at a ratio of 1:4. For example, add 4µl of Protein Staining and Loading Buffer (5X, Odorless) into 16µl of protein sample. 3. Boil the mixture at 95ºC for 5 minutes, which can be extended to 10-15 minutes when necessary to ensure that the genome DNA from tissue or cell samples is broken completely to avoid the appearance of sticky materials. Note: Boiling samples must be carried out at 95ºC. Too high temperature (e.g., 100ºC) or too long time of boiling (e.g., more than 15 minutes) could cause protein degradation or aberrance of the tracking dye. 4. Load samples onto SDS-PAGE wells after cooling to room temperature.5. If protein molecular weight standards are needed, non-prestained protein molecular weight standards without any loading buffers or dissolved in water, saline or PBS can be mixed with an appropriate amount of the InstantView™ SDS-PAGE Protein Staining and Loading Buffer (5X, Odorless) to produce protein molecular weight standards after boiling. 6. After electrophoresis, place the gel absent of glass plates on a UV transilluminator to immediately visualize bands, but if using blue light for the observation, removal of glass plates is not necessary. Note 1: Fluorescent protein signal will decrease gradually as exposure time to UV is increased.Note 2: Proteins in gels can also be stained with Coomassie Brilliant Blue, such as Coomassie Blue Staining Solution ( InstantBlue Coomassie Staining Solution ( SDS-PAGE Protein Staining and Loading Buffer (5X, Odorless).Most prestained and non-prestained protein molecular weight standards are not compatible with the InstantView™ SDS-PAGE Protein Staining and Loading Buffer (5X, Odorless). 2. Stability of the InstantView™ SDS-PAGE Protein Staining and Loading Buffer (5X, Odorless).The InstantView™ SDS-PAGE Protein Staining and Loading Buffer (5X, Odorless) is easily oxidized and should be used within one month after preparation. When stored at room temperature, it should be used within one week. When the staining performance decreased, adding TCEP of no more than 2mM as reducing reagents usually can recover its staining performance. 3. There are precipitates in the InstantView™ Protein Loading Buffer (5X, Odorless) and the InstantView™ SDS-PAGE Protein Staining and Loading Buffer (5X, Odorless) when stored in cold.The InstantView™ Protein Loading Buffer (5X, Odorless) and the InstantView™ SDS-PAGE Protein Staining and Loading Buffer (5X, Odorless) contain SDS that precipitates when stored frozen, but will be resolubilized at room temperature.4. InstantView™ Protein Staining Dye is invisible.The amount of InstantView™ Protein Staining Dye is indeed so little that it could not be seen by naked eyes.5. Do the post-translational modifications of protein affect its staining with InstantView™ Protein Staining and Loading Buffer (5X, Odorless)?No, the post-translational modifications do not have any effect on protein staining. 6. Which wavelength is optimal for the excitation of protein bands run with the InstantView™ SDS-PAGE Protein Staining and Loading Buffer (5X, Odorless)?The optimal wavelength of excitation light is 560nm; however, the following wavelengths can be used: 254, 312, 335, 488 and 520nm. In general, the longer the wavelength of these excitation lights, the better the excitation.7. Is SDS-PAGE Loading Buffer (5X, Odorless) still necessary for protein samples that have been mixed with the InstantView™ SDS-PAGE Protein Staining and Loading Buffer (5X, Odorless)?No, the InstantView™ SDS-PAGE Protein Staining and Loading Buffer (5X, Odorless) acts as both the loading dye and the protein visualization dye.

Specifications

Product Name

UltraBio™ SDS-PAGE Protein Staining and Loading Buffer

Specifications & Purity

5X, Odorless

Molecule Type

Bioreagents/Buffer

Storage and Shipping

Concentration

5X, Odorless

Storage

Protected from light,Store at -20°C

Shipped In

Ice chest + Ice pads

Stability And Storage

Store at -20 ℃ away from light, effective for one year. Stored at 4 ℃ in the dark, effective for 3 months; Stored at room temperature, valid for 2 weeks.