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Pierce Clear Milk Blocking Buffer


Long shelf-life replacement for homemade milk blocking buffers.

Pierce Clear Milk Blocking Buffer

The Thermo Scientific Pierce Clear Milk Blocking Buffer (10X) is a long shelf-life, high performance replacement for homemade milk blocking buffers in Western blotting applications.

The Thermo Scientific Pierce Clear Milk Blocking Buffer (10X) is a proprietary pre-formulated milk solution for blocking excess nonspecific binding sites, reducing background in Western blotting applications and diluting antibodies when used with nitrocellulose and PVDF membranes. Pierce Clear Milk Blocking Buffer (10X) provides lower background, enhanced sensitivity, extended shelf life and reproducible results compared to homemade buffers based on dissolved non-fat dry milk.

Highlights:

  • Excellent stability – stable for 1 year stored at 4°C, unlike typical homemade milk buffers
  • Convenient – concentrated formulation saves storage space and can be diluted easily to obtain optimal blocking results for specific applications
  • Easy to use – no waiting for powdered milk to dissolve with this ready-to-dilute solution
  • Popular – nonfat milk has been used for many years in a variety of protein methods, although it is not recommended for avidin-based techniques because it contains some endogenous biotin

Product Details:

Clear Milk Blocking Buffer for chromogenic and chemiluminescent Western blot detection
Milk blocking buffer compatibility with TMB and ECL substrate detection.Serial dilutions of HeLa cell lysate (7.5, 3.45, 1.88, 0.94, 0.47, 0.23 and 0.12μg) were prepared and separated by electrophoresis. The proteins were transferred to nitrocellulose membranes (Part No. 88018) and the membranes were blocked for 45 minutes with 5% skim milk in TBS + 0.05% Tween™ 20, Clear Milk Blocking Buffer 1X in ultrapure water (Part No. 37587) or StartingBlock T20 (PBS) Blocking Buffer (Part No. 37539). After blocking, the membranes were incubated with suggested concentrations of primary antibodies to targets A, B and C. The membranes were washed and then incubated with of HRP-Conjugated Goat Anti-Mouse IgG (Part No. 31430) and HRP-Conjugated Goat Anti-Rabbit IgG (Part No. 31460) (both at 0.067μg/mL) and then washed again. Working solution of Pierce ECL Substrate (Part No. 32106) was prepared according to the instructions and added to once set of membranes for 5 minutes. The membranes were removed from the substrate, placed in plastic sheet protectors and exposed to CL-XPosure Film (Part No. 34090) for 5 seconds. The other set of membranes were placed in 10mL of Pierce 1-Step Ultra TMB-Blotting Solution (Part No. 37574) and the color development was stopped at 5 minutes by rinsing the membranes with water.

Antibodies:
A. Heat Shock Protein 86 (Hsp86) (Part No. PA3-013)
B. Actin (Part No. MA1-744)
C. Cyclophilin B (Part No. PA1-027A)

 

Clear Milk Blocking Buffer compatibility with nitrocellulose and PVDF membranes
Milk blocking buffer compatibility with nitrocellulose and PVDF membranes. Serial dilutions of A549 cell lysate (5, 2.5, 1.25, 0.62, 0.31, 0.16 and 0.78μg) were prepared and separated by electrophoresis. The proteins were transferred to nitrocellulose (Part No. 88014) and PVDF membranes (Part No. 88585) and the membranes were blocked for 30 minutes with 5% skim milk in PBS + 0.05% Tween™ 20 or Clear Milk Blocking Buffer (Part No. 37587) prepared at 1X in ultrapure water. After blocking, the membranes were incubated with primary antibodies to targets A, B and C. The membranes were washed and then incubated with 10ng/mL of HRP-Conjugated Goat Anti-Mouse IgG (Part No. 31430) and 6.7ng/mL of HRP-Conjugated Goat Anti-Rabbit IgG (Part No. 31460) then washed again. Working solution of SuperSignal West Dura Extended Duration Substrate (Part No. 34076) was prepared according to the instructions and added to the membranes for 5 minutes. The membranes were removed from the substrate, placed in plastic sheet protectors and exposed to CL-XPosure Film (Part No. 34090) for 5 seconds.

Antibodies:
A. Heat Shock Protein 86 (Hsp86) (Part No. PA3-013) at 1μg/mL
B. NFκB (Millipore No. 06-418; 1mg/mL) at 1μg/mL
C. PP2A (Millipore No. 05-421) at 0.2μg/mL

Related Resources:

Blocking buffers for Western blot and ELISA -- technical guide
Blocking strategies for immunohistochemistry -- technical guide

Related Products:

Substrates for Western blotting
Membranes for Western blotting
Stripping buffers for Western blotting
Other blocking buffers


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
37587 Clear Milk Blocking Buffer (10X)
Formulation: Proprietary pre-formulated milk solution
Sufficient For: 30 mini blots at 30mL each of 1X buffer
100mL Product Instructions for product #37587 Clear Milk Blocking Buffer (10X) MSDS for product #37587 Clear Milk Blocking Buffer (10X) Certificate of Analysis for product #37587 Clear Milk Blocking Buffer (10X) $128.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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