ChonBlock Blocking/Sample Dilution Buffer

Properties

Form: solution

Quality level: 100,

Use

  • enough for 10 blots (11921673001 [100 cm2])
  • enough for 60 transfers (11921681001 [100 cm2])

Packaging

  • 100 mL package (11921673001)
  • 6 × 100 mL package (11921681001)

Manufacturer / trade name: Roche

Sent in: wet ice

Storage temperature: 2-8 ° C

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 membrane blocking, membrane washing steps, and dilution of detection antibodies.

Quality

Each lot is function tested (dot blot).

Physical form

The Western Blocking Reagent is a 10x solution containing 10% purified casein protein in the maleic acid buffer.

Preparation note

Working Solution: Preparing Working Solutions

  • Tris-buffered saline (TBS), pH 7.5

At 800 ml redist. water add 6.05 g of Tris base (50 mM), 8.76 g of sodium chloride (150 mM) and adjust the pH to 7.5 with approx. 9.5 ml of 1 M hydrochloric acid. Then dilute to 1 l with double distillation. Water. TBS is stable for 3 months when stored between 2 and 8 ° C.

Note: Since sodium azide inhibits POD, it should not be used as an antimicrobial agent when using POD conjugates.

  • TBS-Tween (TBST)

Wash buffer 1: dissolve 1 ml of Tween 20 in 1 µl of TBS. TBST is stable for 3 months when stored at 2-8 ° C.

Note: 0.1% Tween 20 is suitable for most applications, but depending on the membrane and antibody used, different detergents (such as SDS, Triton X-100 and Nonidet P40 and detergent concentrations of 0, 01 to 1% can lead to better results.

  • Blocking solution (1%)

Dilute 10 ml of Western Blocking Reagent (10x conc.) In 90 ml of TBS. The blocking solution can be stored at 2-8 ° C for 1 month.

Note: Since sodium azide inhibits POD, it should not be used as an antimicrobial agent when using POD conjugates.

  • Blocking solution (0.5%)

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

  • Antibody solutions

The dilution and incubation solution for all antibodies is 0.5% Blocking Reagent in TBS (see above). To take advantage of the system’s full detection potential, we recommend optimizing the primary and secondary antibody dilutions in advance in dot blot assays. (Start with 3 to 4 dilutions of primary antibody first and a constant concentration of the second antibody. Then choose the most suitable dilution of the primary antibody and optimize the secondary antibody concentration in the same way).

Note: The concentration of the blocking reagent is an important parameter to improve the signal-to-noise ratio in Western blots. If a high background appears even at optimized antibody concentrations, increase the Blocking Reagent concentration during antibody incubations and wash steps from 0.5% to 1%. For weak signals, even with prolonged antibody incubations, reduce the concentration of blocking reagent during antibody incubations and wash steps from 0.5% to 0.1%.

Other notes

For life science research only. It should not be used in diagnostic procedures.

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