missing translation for 'onlineSavingsMsg'
Learn More
  1. Home
  2. Products
  3. Western Blot Products
  4. General Western Blot Reagents
  5. Thermo Scientific™ Restore™ Fluorescent Western Blot Stripping Buffer

Thermo Scientific™ Restore™ Fluorescent Western Blot Stripping Buffer

Catalog No. FER62300X4
Change view
Click to view available options
Quantity:
20 mL
100 mL
4 x 100 mL
3 product options available for selection
Product selection table with 3 available options. Use arrow keys to navigate and Enter or Space to select.
Catalog No. Quantity
FER62300X4 4 x 100 mL
PI62299 20 mL
PI62300 100 mL
Use arrow keys to navigate between rows. Press Enter or Space to select a product option. 3 options available.
3 options
Catalog No. FER62300X4 Supplier Thermo Scientific™ Supplier No. 62300X4
Only null left

Description

Quickly and gently remove primary and near-infrared (IR) dye-labeled secondary antibodies from Western blots.

Thermo Scientific™ Restore Fluorescent Western Blot Stripping Buffer is a gentle and highly effective reagent for quickly removing primary and near-infrared (IR) dye-labeled secondary antibodies from Western blots.

Restore Fluorescent Western Blot Stripping Buffer enables the reuse of PVDF membranes, simplifying the Western blot optimization process and allowing the same blot to be reprobed with different primary antibodies to detect alternative targets. Restore Fluorescent Western Blot Stripping Buffer is for use with low-fluorescence PVDF membrane only (Part No. 22860).

Highlights:

  • Fast – strip blots in only 15 minutes at room temperature

  • Saves time – no need to run new gels and prepare a new blot

  • Conserve samples – reprobe the same PVDF membrane for multiple targets

  • Economical – less expensive than other commercially available stripping buffers

  • Efficient – effectively strips blots the first time

Specifications

Form Liquid
Product Line Restore
Quantity 4 x 100 mL
Content And Storage Upon receipt store at room temperature.

Frequently Asked Questions (FAQs)

Can I strip membranes with Restore or Restore Plus Western Blot Stripping Buffer and reprobe for subsequent detection with fluorescent probes?

No. Although the fluorescent antibodies, like other antibodies, can be stripped with Restore Buffers, stripped membranes typically produce unacceptable background for subsequent fluorescent detection methods. We recommend Restore Fluorescent Western Blot Stripping Buffer, Cat. No. 62299 (20 mL) and Cat. No. 62300 (100 mL).

Note: Restore Fluorescent Western Blot Stripping Buffer is for use with low-fluorescence PVDF membrane (Cat. No. 22860) only.

How can I store, strip, and reuse my western blot?

For nitrocellulose or PVDF membrane following Western blot detection using a chemiluminescent or fluorescent substrate system: Following transfer, air dry the membrane and place in an envelope, preferably on top of a supported surface to keep the membrane flat. The blot can be stored indefinitely at -80 degrees C. When ready to reprobe, prewet the PVDF blot with alcohol for a few seconds, followed by a few rinses with pure water to reduce the alcohol concentration. Then proceed as normal with blocking step.

FOR STRIPPING/REPROBING OF MEMBRANES: Harsh protocol (see NOTE below for modifications)

1) Submerge the membrane in stripping buffer (100 mM BME, 2% SDS, 62.5 mM Tris-HCl, pH 6.7) and incubate at 50 degrees C for 30 min with occasional agitation. If more stringent conditions necessary, incubate at 70 degrees C.

2) Wash 2 x 10 min in TBS-T/PBS-T at room temperature.

3) Block the membrane by immersing in 5% blocking reagent TBS-T or PBS-T for 1 hr at room temperature.

4) Immunodetection

NOTE: Often you don't need such harsh conditions to remove antibodies from their proteins. The stringency of one or several of the variables can be decreased: lower the temperature, decrease the time, less BME, less SDS, etc. An especially mild but still often effective stripping protocol is lower pH incubation. Example: pH 2.0 Tris 50-100 mM, 30-60 min incubation (you may do two incubations if you wish). Then rinse and block as usual. If you do not wish to re-use the membrane immediately after stripping, you can store the membrane in plastic wrap (wet, you do not want it to dry out). Another simple, mild stripping buffer is 0.1 M glycine•HCl (pH 2.5-3.0), incubation 30 min to 2 hrs room temperature or 37 degrees C, depending on the antibody.

For Research Use Only. Not for use in diagnostic procedures.