ESO wikiadmin at 13:42, 27 February 2025

2025-02-27T13:42:27Z

← Older revision		Revision as of 15:42, 27 February 2025
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			\| colspan="2" \|⚠️ for stripping use the old, already blue stained boxes ⚠️
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ESO wikiadmin: Created page with "{| class="wikitable" |+ !Materials ! |- |Mild Stripping Buffer |15 g Glycin, 1 g SDS, 10 ml Tween20 in 1L ddH₂O pH2.2 |- |Harsh Stripping Buffer |40 ml Stacking gel buffer (0,5M Tirs/HCl, pH6,8) 10 ml SDS 20%, 200 ml ddH₂O |- | |1X PBS |- | |TBST |} == Procedure mild stripping == * Remove ECL from membrane by washing membrane briefly with ddH₂O * Wash membrane 2x in Mild stripping buffer for 10 min * Wash membrane 2x in PBS for 10 min *..."

2025-02-27T13:40:52Z

Created page with "{| class="wikitable" |+ !Materials ! |- |Mild Stripping Buffer |15 g Glycin, 1 g SDS, 10 ml Tween20 in 1L ddH2O pH2.2 |- |Harsh Stripping Buffer |40 ml Stacking gel buffer (0,5M Tirs/HCl, pH6,8) 10 ml SDS 20%, 200 ml ddH2O |- | |1X PBS |- | |TBST |} == Procedure mild stripping == * Remove ECL from membrane by washing membrane briefly with ddH2O * Wash membrane 2x in Mild stripping buffer for 10 min * Wash membrane 2x in PBS for 10 min *..."

New page

{| class="wikitable"
|+
!Materials
!
|-
|Mild Stripping Buffer
|15 g Glycin, 1 g SDS, 10 ml Tween20 in 1L
ddH2O
pH2.2
|-
|Harsh Stripping Buffer
|40 ml Stacking gel buffer (0,5M Tirs/HCl, pH6,8)
10 ml SDS 20%, 200 ml ddH2O
|-
|
|1X PBS
|-
|
|TBST
|}

== Procedure mild stripping ==

* Remove ECL from membrane by washing membrane briefly with ddH2O
* Wash membrane 2x in Mild stripping buffer for 10 min
* Wash membrane 2x in PBS for 10 min
* Wash membrane 2x for 5 min in TBS-T
* Block in Blotto for 1h at RT
* Apply 1st AB overnight, at 4°C

== Procedure harsh stripping ==

* Prewarm water bath to 50°C
* Remove ECL from membrane by washing membrane briefly with ddH2O
* Add 10 ml of Harsh stripping buffer and supplement with 70 µl β- Mercaptoethanol
* Incubate the membrane for 20 min at 50°C, while shaking
* Wash the membrane under running tap water
* Wash for 1h with ddH2O, change ddH2O every 10 min
* Wash for 1h with TBS-T, change TBS-T every 10 min
* Block in Blocking buffer (Blotto) at least for 1h at RT
* now the membrane is ready for another round of probing

Western Blot: stripping of the membrane - Revision history

ESO wikiadmin at 13:42, 27 February 2025