UltraBio™ Blocking Buffer(10X)

Cat. No.: B751615

Descripción general

UltraBio™ Blocking Buffer from Aladdin is a new generation of rapid and highly effective blocking buffer. Its overall effect is significantly better than that of traditional blocking buffers based on BSA (bovine serum albumin), skim milk powder, casein and the like, as well as similar products from abroad. It can be used for blocking in Western blot (WB), immunofluorescence (IF), immunohistochemistry (IHC), immunocytochemitry (IC) and other experiments, primary or secondary antibody dilution. This product is a 10-fold concentrated (10X) solution that does not contain a detergent. UltraBio™ blocking solution is fast and efficient. The blocking time is usually only 5-15 minutes, and compared with traditional blocking solutions such as BSA, skim milk powder, casein, and similar fast blocking solutions from abroad, it shows a stronger signal-to-noise ratio (see Figure 1). UltraBio™ blocking solution has a very low background after blocking. The blocking solution does not contain serum or albumin, ensuring an extremely high signal-to-noise ratio. UltraBio™ blocking solution is compatible with horseradish peroxidase (HRP), alkaline phosphatase and biotin-labeled secondary antibodies. This product contains a preservative that does not affect the activity of HRP and AP, and will not interfere with the detection of HRP- or AP-labeled secondary antibodies. It also does not contain biotin, which will not interfere with biotin-based detection. UltraBio™ Blocking Solution (10X) is flexible to use. This product is a 10X solution that can be diluted to 1X with PBS, TBS or other appropriate solution according to experimental needs and then used in related experiments. It can also be used for blocking, primary antibody or secondary antibody dilution by adding Tween-20 or Triton X-100 to a final concentration of 0.05%-0.1% according to experimental needs. The blocking effect of this product compared with BSA and similar foreign products is shown in Figure 1. Under the same sample and experimental conditions, only when there are differences in the blocking solution and blocking time as shown in the figure below, the overall background after blocking with UltraBio™ blocking solution from Aladdin is significantly lower than that after blocking with BSA, and the brightness of the target band is significantly higher than that of similar foreign brands. Figure 1. Comparison of the blocking effect of UltraBio™ blocking solution with BSA and similar foreign products. From left to right in each group of experiments: 5 μl protein marker, 2.5 μg protein amount of HeLa cell lysate, 5 μg protein amount of HeLa cell lysate. Please note that for Antibody A, UltraBio™ blocking solution has a lower background than BSA and similar foreign products, and the signal is significantly stronger than similar foreign products. For Antibody B, UltraBio™ Blocking Buffer has lower background than BSA and a significantly stronger signal than foreign similar products. Actual experimental results may vary due to differences in samples, antibodies, experimental conditions, etc. The data in the figure is for reference only. For a comparison and selection of different blocking solutions, please refer to the relevant Aladdin webpage: http://www.aladdin-e.com/. Based on the calculation that 5-10 ml of UltraBio™ blocking solution (1X) is required to block each membrane, one package of this product can block 100-200 membranes.

Note:

This product is recommended for single use only. Reuse may result in a decrease in blocking efficiency. However, for some primary antibodies with a high signal-to-noise ratio, such as some internal reference antibodies, this blocking solution can be reused 2-3 times. Do not mix the recovered blocking solution with unused blocking solution. Usually, this product is used for PVDF membranes and NC membranes with a blocking time of 5-15 minutes. For some antibodies with very high backgrounds, you can try to extend the blocking time to 30-60 minutes. In addition, if there is a special need, it is completely fine to block overnight at 4 °C. Since no blocking solution is suitable for all experimental systems, for some special experiments, it may be necessary to consider using other more suitable blocking solutions based on specific circumstances. Flat-tipped forceps should be used to pick up and place the PVDF membrane and NC membrane, and only the edges should be gently clamped. During the operation, scratches, creases or indentations on the membrane surface should be avoided. Once the PVDF membrane is wetted and activated, it needs to be kept wet at all times. According to the specific steps of Western, it can be placed in an appropriate solution such as a western transfer solution or washing solution. Otherwise, an abnormal background that is difficult to seal may be generated. In the case of high background, to further improve the signal-to-noise ratio, this product can be diluted to 1X and then Tween-20 or Triton X-100 can be added to a final concentration of 0.05%-0.1%. This product is for professional scientific research use only, and is not intended for clinical diagnosis or treatment, nor for use in food or drugs. For your safety and health, please wear laboratory clothing and disposable gloves when handling.

Instructions for use:

1. Dilution of UltraBio™ Blocking Buffer (10X) Dilute UltraBio™ Blocking Buffer (10X) to 1X with PBS, TBS or PBS or TBS containing 0.05%-0.1% Tween-20 or Triton X-100 according to the needs of the experiment.

2. Blocking of membranes in Western Blot

a. After the transfer is complete, wash the protein membrane with Western washing solution for 1-2 minutes.

b. Depending on the size of the membrane, pour a certain volume of UltraBio™ blocking solution (1X) into a petri dish or other appropriate container to ensure that the blocking solution can fully cover the membrane later. For a conventional western, it is recommended to use about 10 ml of blocking solution for a membrane about 6.6 × 8.5 cm.

c. Use flat-tipped forceps to hold one corner of the membrane, place the membrane in UltraBio™ blocking solution (1X), completely submerge the membrane in the solution, and place it on a horizontal shaker to block for about 10 minutes (usually 5-15 minutes is fine; testing with multiple antibodies has shown that the effect of 10 minutes of blocking is often significantly better than the effect of the conventional 1-hour BSA blocking).

d. The blocked membrane can then be used for subsequent experiments such as primary antibody incubation.

3. For IF, IHC and other experiments, simply follow the relevant experimental procedures and directly replace the traditional blocking solution with UltraBio™ blocking solution (1X). The general blocking time can be shortened to 10 minutes (among the multiple primary antibodies tested by Aladdin, there is no significant difference in the effect of blocking for 10-20 minutes, and the effect of blocking for 10 minutes is equivalent to or significantly better than the conventional blocking method).

Specifications

Product Name

UltraBio™ Blocking Buffer(10X)

Tipo de molécula

Bioreagents/Buffer

Almacenamiento y envío

Condiciones de almacenamiento de almacenamiento

Store at 2-8°C

Enviado en

Wet ice

Estabilidad y almacenamiento

Store at 4℃ for up to 1 year, or -20℃ for long-term storage. The 1X dilution should be kept at 4℃ and use it within 1 month.