All lanes : Anti-Nicotinic Acetylcholine Receptor alpha 5/CHRNA5 antibody [EPR24135-98] (ab259859) at 1/1000 dilution

Lane 1 : Human Hippocampus tissue lysate
Lane 2 : Human brain tissue lysate
Lane 3 : Human testis tissue lysate
Lane 4 : Human kidney tissue lysate
Lane 5 : Human liver tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution (VeriBlot for IP secondary antibody(HRP))

Predicted band size: 53 kDa
Observed band size: 53 kDa

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

The observed MW is consistent with what has been described in the literature (PMID:30543688).

Low expression: kidney, liver (PMID:20920278).

Exposure time: Lane 1: 10 seconds

Lane 2-5: 26 seconds

 

All lanes : Anti-Nicotinic Acetylcholine Receptor alpha 5/CHRNA5 antibody [EPR24135-98] (ab259859) at 1/1000 dilution

Lane 1 : A549 (human lung carcinoma epithelial cell) whole cell lysate
Lane 2 : MCF7 (human breast adenocarcinoma epithelial cell) 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: 53 kDa
Observed band size: 53 kDa

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

The observed MW and expression pattern is consistent with what has been described in the literature (PMID:30543688, PMID:28878681).

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

Exposure time: Lane 1: 3 minutes

Lane 2: 26 seconds

 

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized A549 cells labelling Nicotinic Acetylcholine Receptor alpha 5/CHRNA5 with ab259859 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in A549 cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

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

All lanes : Anti-Nicotinic Acetylcholine Receptor alpha 5/CHRNA5 antibody [EPR24135-98] (ab259859) at 1/1000 dilution

Lane 1 : Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate
Lane 2 : Mouse testis tissue lysate
Lane 3 : Mouse kidney tissue lysate
Lane 4 : Mouse liver 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: 53 kDa
Observed band size: 53 kDa

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

The observed MW is consistent with what has been described in the literature (PMID:30543688).

Low expression: kidney, liver (PMID:20920278).

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

Exposure time: 3 minutes

 

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neuron cells labelling Nicotinic Acetylcholine Receptor alpha 5/CHRNA5 with ab259859 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing positive staining in mouse primary neuron. Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection is observed. ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

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

All lanes : Anti-Nicotinic Acetylcholine Receptor alpha 5/CHRNA5 antibody [EPR24135-98] (ab259859) at 1/1000 dilution

Lane 1 : Rat hippocampus tissue lysate
Lane 2 : Rat brain tissue lysate
Lane 3 : Rat testis tissue lysate
Lane 4 : Rat kidney 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: 53 kDa
Observed band size: 53 kDa

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

The observed MW is consistent with what has been described in the literature (PMID:30543688).

Low expression: kidney (PMID:20920278).

Samples (except lane1) are non-boiled as boiling may cause protein aggregates.

Exposure time: 26 seconds

 

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary neuron cells labelling Nicotinic Acetylcholine Receptor alpha 5/CHRNA5 with ab259859 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing positive staining in rat primary neuron. Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection is observed. ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

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