Western blot membrane stripping for restaining protocol

antibody stripping
antibody stripping

Our protocol for western blot membrane stripping and restaining contains step-by-step particulars on the elimination of antibodies from western blot membranes.

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Stripping is the time period used to explain the elimination of major and secondary antibodies from a western blot membrane. Stripping is beneficial when one needs to research a couple of protein on the identical blot, as an illustration a protein of curiosity and a loading management. When probing for a number of targets, stripping and re-probing a single membrane as a substitute of working and blotting a number of gels has the benefit of saving samples, supplies, and time.

It’s not advisable to make quantitative comparisons of targets probed earlier than and after stripping because the process removes some pattern protein from the membrane. For a similar purpose, a stripped membrane shouldn’t be probed to reveal the absence of a protein.

 

The next two protocols differ in harshness of therapy. As a rule of thumb, attempt the gentler one first after which proceed to the harsher one if there may be nonetheless a sign from the antibody that one is making an attempt to strip. These steps will be repeated for probing with a number of antibodies, although the potential sign could also be weaker and the background increased after every spherical of stripping. Some researchers report efficiently staining a membrane after stripping ten or extra instances.


antibody stripping buffer
antibody stripping buffer

Effectivity of stripping will be checked by incubating the membrane with chemiluminescent detection reagent. If stripping is judged to be passable, rinse the membrane a number of instances with buffer, then block earlier than continuing to the antibody incubation.

Stripping and Reprobing Western Blotting Membranes

Western blotting is a generally used approach for learning protein operate and localization. Sometimes, protein samples are separated by SDS-PAGE and transferred to a nitrocellulose or PVDF membrane which is then probed with particular antibodies. In contrast to nucleic acid based mostly applied sciences, which permit reuse of Southern and Northern blots, it has been troublesome to reuse Western blots.

Stripping and re-probing of Western blots gives a number of benefits:

  1. Conservation of samples which might be costly or accessible solely in restricted portions;
  2. Evaluation of a given blot utilizing a number of totally different antibodies, e.g. subtype- or isoform-specific antibodies;
  3. Re-analysis of anomalous outcomes and affirmation with the identical or a unique antibody;
  4. Correcting errors in incubation with the incorrect antibody;
  5. Price financial savings in reagents and time by reusing the identical blot.

 

ECL Plus Western Blotting Substrate

AR1196 50mL(sufficient reagents for 500 cm2 of membrane)
EUR 101

ECL Western Blotting Substrate Kit

K820-50
EUR 153

ECL Western Blotting Substrate Kit

K820-500
EUR 338

ECL Western Blotting Substrate Kit

K2187-50 50 assays
EUR 160

ECL Western Blotting Substrate Kit

K2187-500 500 assays
EUR 363

WESTSAVE Up (Western Blotting Substrate)

LF-QC0101 1 kit
EUR 151
Description: Abfrontier WESTSAVE UpTM (Western Blotting Substrate) is enhanced luminol-based chemiluminescent substrate for the non-radioactive detection of Horseradish Peroxidase(HRP) labeled antibodies.

WESTSAVE Up (Western Blotting Substrate, double pack)

LF-QC0102 1 kit(Double pack)
EUR 222
Description: Abfrontier WESTSAVE UpTM (Western Blotting Substrate) is enhanced luminol-based chemiluminescent substrate for the non-radioactive detection of Horseradish Peroxidase(HRP) labeled antibodies.

Western Blotting Filter Paper, 12.5 cm—12.5 cm

AR0172 100 sheets
EUR 70

Western Blotting Filter Paper, 9 cm— 7.5 cm

AR0173 100 sheets
EUR 65

Purified Lassa fever Nucleoprotein control for Western Blotting

LFNP15-C-10 100 ul
EUR 286

Purified Nipah virus Glycoprotein control for Western Blotting

NIV11-C 100 ul
EUR 286

Purified Nipah virus Nucleoprotein control for Western Blotting

NIV21-C 100 ul
EUR 225

Femto ECL substrate for ELISA/Western Blotting (2 Components)

80210 100 ml
EUR 347

GloBrite chemiluminescence reagent kit for western blotting 200 mL

GLB1 200 mL
EUR 145

GloBrite chemiluminescence reagent kit for western blotting 500 mL

GLB2 500 mL
EUR 234

Mayaro virus (MAYV) Capsid Protein control for Western Blotting

MAYV31-C 100 ul
EUR 286

Mayaro virus (MAYV) nsP1 protein control for Western Blotting

MAYV41-C 100 ul
EUR 286

Mayaro virus (MAYV) 6K Protein control for Western Blotting

MAYV51-C 100 ug
EUR 286

Mayaro virus (MAYV) E3 protein control for Western Blotting

MAYV61-C 100 ul
EUR 286

Western Blotting Mouse IgG DAB Chromogenic Reagent Kit (Yellow)

SA2020 1 kit(1200 cm2)
EUR 141

Western Blotting Goat IgG DAB Chromogenic Reagent Kit (Yellow)

SA2021 1 kit(1200 cm2)
EUR 141

Western Blotting Rabbit IgG DAB Chromogenic Reagent Kit (Yellow)

SA2022 1 kit(1200 cm2)
EUR 141

Western Blotting Rat IgG DAB Chromogenic Reagent Kit (Yellow)

SA2023 1 kit(1200 cm2)
EUR 141

Western Blotting Mouse IgG DAB Chromogenic Reagent Kit (Blue)

SA2024 1 kit(1200 cm2)
EUR 141

Western Blotting Rabbit IgG DAB Chromogenic Reagent Kit (Blue)

SA2025 1 kit(1200 cm2)
EUR 141

Blotting roller

2322479 4unit
EUR 267

Ribulose-1,5-bisphosphate carboxylase oxygenase (Rubisco/Rbcl) Loading Control for Western Blotting

RBCL15-C 100 ul
EUR 286

Western Blotting Boxes, 6-Compartment, 1 3/8 Height ( 32x100 mm Each component)

LC7012-004 4/Pk
EUR 97

Western Blotting Boxes,15.2 x 10.2 x 5.1 cm (6 x 4 x 2in)

LC7021-005 5/Pk
EUR 85

Western Blotting Boxes, 15.2 x 10.2 x 3.2 cm (6 x 4 x 1 1/4in)

LC7020-005 5/Pk
EUR 84

Blotting Paper, 10X15cm, 100 Sheets

PP3322 1PK, 100UNIT
EUR 105.68
  • Product category: Electrophoresis Related/Blotting Papers

Blotting Paper, 15X15cm, 100 Sheets

PP3323 1PK, 100UNIT
EUR 117.04
  • Product category: Electrophoresis Related/Blotting Papers

Blotting Paper, 20X20cm, 100 Sheets

PP3324 1PK, 100UNIT
EUR 166.4
  • Product category: Electrophoresis Related/Blotting Papers

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