Fluorescent protein loading buffer is used to pre-stain SDS-PAGE protein samples at the sample treatment stage before electrophoresis and fluorescent label is applied. After the experiment is completed, the protein strips in the gel or after the transfer film can be directly observed and analyzed by ultraviolet lamps, LED lamps or other digital imaging systems, without dyeing and decolorization. Fluorescent protein loading buffer is more sensitive than silver dye. Strong stability, low background. All proteins can be stained without affecting protein mobility and electrophoretic profile. During the electrophoresis process, the free dye molecules migrated at the same rate as bromophenol blue, and moved to the end of the gel at the end of the gel running, and the background was clean. Fluorescent protein loading buffers are ideal for SDS-PAGE for protein expression tracking and purification as well as Western blot.

Usage steps

1. Add the resuspension buffer according to the volume on the tube wall label before use. Resuspension buffer precipitation may occur at 4℃, and let it sit at room temperature until the precipitation disappears.

2. Mix the protein sample treated with the sample buffer with the protein sample to be electrophoreted 1:2. For example, 3ul loading buffer + 6ul protein sample.

3. Heat the sample mixture at 90-100℃ for 5 minutes. For cell or tissue samples, the heating time is extended to 10 minutes. Ensure heating temperature > The sample is fully heated at 90℃. (See Note 3)

4. The sample can be loaded for electrophoresis (no Loading Buffer is required). After electrophoresis, the gel is placed on the transmissometer for observation and photography. The transmissometer can be an ultraviolet lamp, a blue LED lamp or other gel imaging system. If the light source is in the visible wavelength category, there is no need to peel the glue, because the light in the visible wavelength range can penetrate the plastic plate made of glass or plastic.

6. (Optional) If necessary, gels treated with fluorescent protein loading buffer can still be tested. Follow the standard test procedure.

Note:

1. Do not use the loading buffer to treat the molecular weight standard of pre-dyed or pre-treated protein. These products are not compatible with fluorescent protein loading buffers.

2. Sensitivity influencing factors: high pH (greater than 7) will not affect the dyeing, low pH will reduce the efficiency of dyeing, if the pH of the sample is lower than 5, it is recommended to adjust the pH above 7.

3. Most prefabricated adhesives (except Bis-Tris system) can proceed directly to the next step. Bis-Tris preforms, such as Life Technology, need to add an Enhancing buffer of 20% of the treated sample volume. For example, 2ul Enhancing buffer is added to a 10ul processed sample.

4. Fluorescent protein loading buffer is not suitable for use in SDS-PAGE experiments such as cutting protein bands for sequencing, mass spectrometry or antibody preparation.

5. It is recommended to store at -20℃, if it is left at room temperature for 2-3 weeks, new DTT can be added, and the protein strip will become clear and bright again. The final concentration of DTT added should not exceed 20 mM. Other reducing agent TCEP(2 mM) can also be added, and the effect is also very good.

Note：Product information may be optimized and upgraded. Please refer to the actual label information for accuracy.