Anti-EAAT1 antibody (ab416)

  Western blot - EAAT1 antibody (ab416)

Anti-EAAT1 antibody (ab416) + Rat brain cortex

Predicted band size : 60 kDa
Observed band size : 60 kDa
Additional bands at : 150 kDa (possible dimer).

  Immunohistochemistry (PFA perfusion fixed frozen sections) - EAAT1 antibody (ab416)

ab416 at 1/100 dilution staining EAAT1 in mouse coronal tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Adult Sprague–Dawley rats were injected intraperitoneally with pentobarbitone and were transcardially perfused-fixed with heparinized saline and 4% paraformaldehyde in PBS. Brains were removed, were postfixed in 4% paraformaldehyde for 5h, and were stored in PBS. Sections of each brain were pretreated with citrate buffer for 30 min at 65°C to increase antigen retrieval and penetration of the antibodies into the tissues. Sections were permeabilized with 1% Triton X-100 for 5 min and blocked with 3% normal horse serum in 0.1 M PBS, pH 7.4, for 60 min and incubated at room temperature for 48h with primary antibodies. An Alexa Fluor® 488 conjugated anti rabbit was used as secondary at 1/1000 dilution.

Image from Barcia C. et. al., PLoS One. 2008 Aug 20;3(8):e2977 (Fig 8C).

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - EAAT1 antibody (ab416)

ab416 (1:500) staining EAAT1 in human cerebellum using an automated system (DAKO Autostainer Plus). Using this protocol there is strong staining of membrane cells in the purkinje glial region .
Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.