Anti-GNAO1 antibody [EPR24151-69] (ab259884) at 1/1000 dilution + Human cerebellum tissue lysate at 10 µg

Secondary
VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/5000 dilution (VeriBlot for IP secondary antibody(HRP))

Predicted band size: 40 kDa

Blocking and diluting buffer and concentration: 5% NFDM/TBST

Exposure time: 5.5 seconds

 

All lanes : Anti-GNAO1 antibody [EPR24151-69] (ab259884) at 1/1000 dilution

Lane 1 : IMR-32 (human neuroblastoma neuroblast) whole cell lysate
Lane 2 : Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate
Lane 3 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Predicted band size: 40 kDa

Blocking and diluting buffer and concentration: 5% NFDM/TBST

Samples are non-boiled as boiling may cause protein aggregates.

Exposure time: 3 minutes

 

All lanes : Anti-GNAO1 antibody [EPR24151-69] (ab259884) at 1/1000 dilution

Lane 1 : Mouse brain tissue lysate
Lane 2 : Mouse cerebellum tissue lysate
Lane 3 : Mouse hippocampus tissue lysate
Lane 4 : Mouse heart tissue lysate
Lane 5 : Rat brain tissue lysate
Lane 6 : Rat heart tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Predicted band size: 40 kDa

Blocking and diluting buffer and concentration: 5% NFDM/TBST

This blot was developed using a higher sensitivity ECL.

Samples are non-boiled as boiling may cause protein aggregates.

 

The MW observed is consistent with the literature (PMID: 14519434).

 Exposure time: 7.75 seconds

 

Immunohistochemical analysis of paraffin-embedded Human cerebellum tissue labelling GNAO1 with ab259884 at 1/2000 (0.261 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human cerebellum. The section was incubated with ab259884 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND RX instrument Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Immunohistochemical analysis of paraffin-embedded Mouse cerebellum tissue labelling GNAO1 with ab259884 at 1/5000 (0.104 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse cerebellum. The section was incubated with ab259884 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND RX instrument Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Immunohistochemical analysis of paraffin-embedded Rat cerebellum tissue labelling GNAO1 with ab259884 at 1/5000 (0.104 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat cerebellum. The section was incubated with ab259884 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND RX instrument Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse cerebellum tissue labeling GNAO1 with ab259884 at 1/100 (5.22 ug/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Positive staining on mouse cerebellum is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat cerebellum tissue labeling GNAO1 with ab259884 at 1/100 (5.22 ug/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Positive staining on rat cerebellum is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).