Western Blot Buffer Kit (Including Blocking Buffer, Wash Buffer (10X), Primary Antibody Dilution Buffer, Secondary Antibody Dilution Buffer)

Cat. No.: W751791

Overview

Aladdin's Western Buffer set includes Blocking Buffer, Wash Buffer, Primary Antibody Dilution Buffer, and Secondary Antibody Dilution Buffer commonly used for Western Blot.Primary Antibody Dilution Buffer for Western Blot included in this product can be used for dilution and preparation of primary antibodies. Its main ingredients including BSA, detergent and primary antibody stabilization reagent, have been optimized to reduce non-specific bindings and stabilize primary antibodies after dilution. The diluted primary antibodies in this product can be stored at 4℃ for six months at least, and can be reused multiple times.Blocking Buffer for Western Blot provided in this product can be used to block PVDF or NC membranes. The main active ingredients including BSA and detergent can effectively block the protein binding sites on membranes and reduce non-specific bindings of antibodies, thus reducing the background and enhancing the signal-to-noise ratio.The Western Wash Buffer (10X) can be used to wash membranes after incubation with antibodies. Proper washing can reduce the background and enhance the signal-to-noise ratio.Secondary Antibody Dilution Buffer for Western Blot can be used to dilute secondary antibodies for Western blot. The main active ingredients including BSA, detergent and secondary antibody stabilization reagent, have been optimized for effective reduction of non-specific bindings and stabilization of secondary antibodies after dilution. The diluted secondary antibodies can be reused 3-5 times within 1-2 weeks when stored at 4℃.

Precautions：

The PVDF and NC membranes should be handled with flat tip tweezers, and only the corners should be gently gripped, avoiding scratches, creases or indentations of the membrane.The activated PVDF membrane needs to be kept moist in the transfer buffer or wash buffer until use. Once accidentally dried, it needs to be re-activated.To recycle antibodies for later use, we recommend incubating membranes in primary antibody solutions at 4℃ if possible to maintain the activity of primary antibodies.Usually, blocking for 60 minutes at room temperature is sufficient. For some antibodies producing high background, block the membrane overnight at 4℃.The Western Wash Buffer (1X) should be stored at 4℃ and used up within 1-2 months, or within 1-2 weeks if stored at room temperature.Secondary Antibody Dilution Buffer for Western Blot is sterilized by filtration, and bacterial contamination should be avoided during use. At room temperature, it should be used up within one month and discarded once contaminated with microorganisms.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. Primary Antibody Dilution Buffer for Western Blota. Dilute the primary antibody appropriately according to manufacture's instructions. The diluted primary antibody in this Primary Antibody Dilution Buffer can be recovered and stored at 4℃ for later use.2. Blocking Buffer for Western Blot a. After protein transfer, wash membranes with the Western Wash Buffer for 1-2 minutes, block membranes for 60 minutes in the Blocking Buffer, then incubate in diluted primary antibody solutions.3. Western Wash Buffer (10X) a. Prepare an appropriate amount of Western Wash Buffer (1X) by diluting the Western Wash Buffer (10X) with distilled water and mix well before use. The diluted Wash Buffer can be stored at room temperature for up to 1-2 weeks, and should be discarded when turned turbid or precipitation occurs.b. After incubation of membranes with primary or secondary antibodies, add sufficient Wash Buffer (1X) to fully cover membranes, and wash for 5 minutes with shaking. Repeat washing for a total of 3 times. Then proceed to the subsequent procedures. If there is high background on membrane, extend the washing time or increase the number of washes appropriately, which usually does not have negative impacts on Western results.4. Secondary Antibody Dilution Buffer for Western Blot a. Dilute the secondary antibody appropriately according to manufacture's instructions. The diluted secondary antibody can be recovered and stored at 4℃ for later use. It can be reused 3-5 times within 1-2 weeks.b. 

Storage and Shipping

Storage

Store at 2-8°C

Shipped In

Wet ice

Stability And Storage

Stored at 4 ℃, valid for one year. Long term inapplicability can be stored at -20 ℃.