Product Information

Features

1. Ultra-fast no-wash blocking: 5 minutes to complete the blocking, no need to wash, directly incubate the primary antibody

2. No protein components: It is very effective in avoiding the appearance of nonspecific bands or high background caused by nonspecific binding of proteins in the blocking solution components with antibodies

3. Commonly used for conventional antibodies, phosphorylated antibodies, and biotin-labeled antibodies

4. Antigen-antibody binding enhancement formula: can effectively increase the antigen-antibody binding ability, better target band signal, and lower background.

Procedure

1. Dilute HRbio™ Rapid Blocking Buffer (10×) to deionized water at a ratio of 1:9 to make 1× Rapid Blocking Buffer working solution;

2. Place the NC or PVDF membrane after transfer into an appropriate volume of 1×HRbio™ rapid blocking solution (note that the membrane must be completely covered. For a 6.5*8.5cm membrane, the recommended amount is 15-20ml), place it on a horizontal shaker, and incubate at room temperature for 5 minutes.

3. Please note: The primary antibody stock solution used must be diluted with 1×HRbio™ Rapid Blocking Solution to the corresponding antibody working solution (the subsequent antibody working solution can be directly recovered and stored at 4 degrees for at least 2 months) to better ensure the low background of subsequent experimental results. The secondary antibody can be diluted normally according to the conventional experimental method.

4. Take out the blocked membrane. Note that there is no need to wash with TBST. Directly put it into the primary antibody working solution prepared in step 2 and incubate normally. After completion, repeat the TBST washing 3 times, each time for 10 minutes.

5. Add the membrane incubated with the primary antibody in step 3 to the secondary antibody working solution prepared in step 2 and incubate. After incubation, wash with TBST three times, 10 minutes each time, and then expose normally.

Storage conditions

Store at 4℃, shelf life is 12 months.

Precautions

1. The blocking solution may not be suitable for some experimental systems. Therefore, for some special experiments or when the antibody effect cannot meet the expectations, you can consider using a more suitable blocking system to conduct the experiment;

2. Weak or no signal:

(1) The amount of target protein sample loaded is too small or the antibody concentration is too low. You can increase the amount of sample loaded or increase the concentration of the primary antibody;

(2) The transfer efficiency is too low, resulting in no target protein or too little target protein on the membrane. After re-transferring, use Ponceau red staining to check that the transfer effect is normal before subsequent blocking and incubation with primary and secondary antibodies;

(3) This is caused by reasons such as low antibody titer and poor specificity, and needs to be adjusted according to the experimental conditions.

3. This product is for scientific research purposes only!

3. High background:

(1) The primary antibody was not diluted with 1×HRbio™ Rapid Blocking Buffer;

Solution: Dilute the primary antibody with 1× HRbio™ Fast Blocking Buffer or HRbio™ Stable Antibody Diluent.

(2) Excessive use of antibodies or excessive protein loading will result in a large number of non-specific bands.

Solution: This can be avoided by reducing the amount of protein loaded or the antibody concentration.

(3) The washing time of the transfer membrane is too few or too short;

Solution: Supplement the number of TBST washes with the time of washing the membrane.

(4) Caused by contamination of reagents or instruments;

Solution: Need to be adjusted according to the specific circumstances of the experiment.