ESO wikiadmin: Created page with "{| class="wikitable" |+ !Materials |- |PBS pH 7.4 |- |Blocking buffer (1% BSA, 1% serum and 0.1% Triton X-100 in PBS) |- |Corresponding 1st and 2nd antibodies |- |Barrier pen |- |Mounting medium |} === Procedure: === * Thaw the slides at room temperature for 10-20 min * The slides are washed for 10 min in PBS * Block non-specific staining by incubating samples in blocking buffer for 30 minutes at RT * Surround the tissue with a hydrophobic barrier using a barrier pen..."

2025-05-20T14:09:55Z

Created page with "{| class="wikitable" |+ !Materials |- |PBS pH 7.4 |- |Blocking buffer (1% BSA, 1% serum and 0.1% Triton X-100 in PBS) |- |Corresponding 1st and 2nd antibodies |- |Barrier pen |- |Mounting medium |} === Procedure: === * Thaw the slides at room temperature for 10-20 min * The slides are washed for 10 min in PBS * Block non-specific staining by incubating samples in blocking buffer for 30 minutes at RT * Surround the tissue with a hydrophobic barrier using a barrier pen..."

New page

{| class="wikitable"
|+
!Materials
|-
|PBS pH 7.4
|-
|Blocking buffer (1% BSA, 1% serum and 0.1% Triton X-100 in
PBS)
|-
|Corresponding 1st and 2nd antibodies
|-
|Barrier pen
|-
|Mounting medium
|}

=== Procedure: ===

* Thaw the slides at room temperature for 10-20 min
* The slides are washed for 10 min in PBS
* Block non-specific staining by incubating samples in blocking buffer for 30 minutes at RT
* Surround the tissue with a hydrophobic barrier using a barrier pen
* Apply primary antibodies diluted in blocking buffer according to manufacturer’s instructions overnight at 4° C (now all incubations are carried out using a humidified chamber)
* The next day slides are washed in PBS 3 times for 15 min
* The slides are incubated in the corresponding 2ary antibody solution in blocking buffer for 1h at RT
* Finally the slides are washed three times in PBS and mounted

Fluorescent Staining of Cryosections - Revision history