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 121.2

WESTSAVE Up (Western Blotting Substrate)

LF-QC0101 1 kit
EUR 181.2
Description: Abfrontier WESTSAVE UpTM (Western Blotting Substrate) is enhanced luminol-based chemiluminescent substrate for the non-radioactive detection of Horseradish Peroxidase(HRP) labeled antibodies.

ECL Western Blotting Substrate Kit

K820-50
EUR 183.6

ECL Western Blotting Substrate Kit

K820-500
EUR 405.6

ECL Western Blotting Substrate Kit

K2187-50 50 assays
EUR 192

ECL Western Blotting Substrate Kit

K2187-500 500 assays
EUR 435.6

ECL PRIME WESTERN BLOTTING SYSTEM

GERPN2232 EACH
EUR 254.22

ECL Prime Western Blotting Det

RPN2232 EACH
EUR 258.78

Amersham ECL DualVue Western Blotting Markers

RPN810 EACH
EUR 223.44

WESTSAVE Up (Western Blotting Substrate, double pack)

LF-QC0102 1 kit(Double pack)
EUR 266.4
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 84

Western Blotting Filter Paper, 9 cm— 7.5 cm

AR0173 100 sheets
EUR 78

Purified Nipah virus Glycoprotein control for Western Blotting

NIV11-C 100 ul
EUR 343.2

Purified Nipah virus Nucleoprotein control for Western Blotting

NIV21-C 100 ul
EUR 270

Purified Lassa fever Nucleoprotein control for Western Blotting

LFNP15-C-10 100 ul
EUR 343.2

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

80210 100 ml
EUR 416.4

Mayaro virus (MAYV) Capsid Protein control for Western Blotting

MAYV31-C 100 ul
EUR 343.2

Mayaro virus (MAYV) nsP1 protein control for Western Blotting

MAYV41-C 100 ul
EUR 343.2

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

MAYV51-C 100 ug
EUR 343.2

Mayaro virus (MAYV) E3 protein control for Western Blotting

MAYV61-C 100 ul
EUR 343.2

GloBrite chemiluminescence reagent kit for western blotting 200 mL

GLB1 200 mL
EUR 174

GloBrite chemiluminescence reagent kit for western blotting 500 mL

GLB2 500 mL
EUR 280.8

Western Blotting Mouse IgG DAB Chromogenic Reagent Kit (Yellow)

SA2020 1 kit(1200 cm2)
EUR 169.2

Western Blotting Goat IgG DAB Chromogenic Reagent Kit (Yellow)

SA2021 1 kit(1200 cm2)
EUR 169.2

Western Blotting Rabbit IgG DAB Chromogenic Reagent Kit (Yellow)

SA2022 1 kit(1200 cm2)
EUR 169.2

Western Blotting Rat IgG DAB Chromogenic Reagent Kit (Yellow)

SA2023 1 kit(1200 cm2)
EUR 169.2

Western Blotting Mouse IgG DAB Chromogenic Reagent Kit (Blue)

SA2024 1 kit(1200 cm2)
EUR 169.2

Western Blotting Rabbit IgG DAB Chromogenic Reagent Kit (Blue)

SA2025 1 kit(1200 cm2)
EUR 169.2

Amersham ECL Western Blotting Detection Reagents for 4000cm2 membrane

RPN2106 EACH
EUR 314.64

Amersham ECL Western Blotting Detection Reagents for 1000cm2 membrane

RPN2109 EACH
EUR 158.46

Amersham ECL Western Blotting Detection Reagents for 6000cm2 membrane

RPN2134 1KIT
EUR 482.22

Amersham ECL Western Blotting Detection Reagents for 2000cm2 membrane

RPN2209 EACH
EUR 245.1

Amersham ECL Western Blotting Systemor detection for 1000 cm2 membrane

RPN2108 EACH
EUR 274.74

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