Merck, 11921681001, Western Blocking Reagent, Solution

Name: Western Blocking Reagent, Solution
Brand: Merck
SKU: 11921681001

Size:6 x 100 mL, blocking reagent for western blots, solution, sufficient for 10 blots (11921673001 [100 cm²]), sufficient for 60 blots (11921681001 [100 cm²])

Catalog Number: 11921681001

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Western Blot Detection Reagents

TagsLabware Labware Partners Roche<sup>®</sup> Products

CollectionsMerck Cell Culture Supplements & Reagents: Complete Solutions for Research & Bioproduction Merck Roche Life Science Products for PCR, RT‑PCR & Nucleic Acid Purification

Description

Application
Western Blocking Reagent is used as a general blocking agent during western blotting experiments and immunofluorescence staining. The reagent is used in steps such as blocking the membrane, membrane washing steps, and diluting detection antibodies.
Physical form
The Western Blocking Reagent is a 10x solution that contains 10% purified casein protein in maleic-acid buffer.
Preparation Note
Working solution: PREPARATION OF WORKING SOLUTIONS

Tris buffered saline (TBS), pH 7.5

To 800 ml redist. water add 6.05 g Tris base (50 mM), 8.76 g sodium chloride (150 mM) and adjust the pH to 7.5 with approx. 9.5 ml 1 M hydrochloric acid. Then dilute to 1 l with double-dist. water. TBS is stable for 3 months, when stored at 2 to 8 °C.
Note: Since sodium azide inhibits POD, it must not be used as antimicrobial agent when using POD-conjugates.

TBS-Tween (TBST)

Washing buffer 1: Dissolve 1 ml Tween 20 in 1 l of TBS. TBST is stable for 3 months, when stored at 2 to 8 °C.
Note: 0.1% Tween 20 is suitable for the most applications, but—depending on the membrane and on the antibody used—different detergents (like SDS, Triton X-100 and Nonidet P40 and detergent concentrations from 0.01 to 1% may lead to better results.

Blocking solution (1%)

Dilute 10 ml Western Blocking Reagent (10x conc.) in 90 ml TBS. The Blocking solution can be stored at 2 to 8 °C for 1 month.
Note: Since sodium azide inhibits POD, it must not be used as antimicrobial agent when using POD-conjugates.

Blocking solution (0.5%)

Dilute 50 ml Blocking solution (1%) with 50 ml of TBS.

Antibody solutions

Dilution and incubation solution for all antibodies is 0.5% Blocking reagent in TBS (see above). In order to exploit the full detection potential of the system we recommend to optimize the dilutions of the primary and secondary antibody in dot blot assays in advance. (Start first with 3 to 4 dilutions of primary antibody and a constant concentration of the second antibody. Then choose the most suitable dilution of primary antibody and optimize the concentration of the secondary antibody in the same way.)
Note: The concentration of the blocking reagent is an important parameter for improvement of the signal to noise ratio in Western blots. If high background appears even under optimized antibody concentrations, increase the concentration of the blocking reagent during the antibody incubations and washing steps from 0.5% to 1%. In case of weak signals even with prolonged antibody incubations lower the concentration of blocking reagent during the antibody incubations and washing steps from 0.5% to 0.1%.
Analysis Note
Each lot is function tested (dot blot).
Other Notes
For life science research only. Not for use in diagnostic procedures.

Specifications

General

UNSPSC Code	12352200
NACRES	NA.54
form	solution
usage	sufficient for 10 blots (11921673001 [100 cm2])
packaging	pkg of 100 mL (11921673001)
manufacturer/tradename	Roche
storage temp.	2-8°C

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Merck, 11921681001, Western Blocking Reagent, Solution

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