Anti-GABA A Receptor gamma 2/GABRG2 antibody [EPR25325-5]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-GABA A Receptor gamma 2/GABRG2 antibody [EPR25325-5] (AB288564)

Flow cytometric analysis of 293T (Human embryonic kidney epithelial cell, Left) / SH-SY5Y (Human neuroblastoma epithelial cell, Right) cells labelling GABA A Receptor gamma 2/GABRG2 with ab288564 at 1/50 dilution (1ug)/ Left and Right. A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Negative control : 293T (PMID : 27864268). Gated on viable cells.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-GABA A Receptor gamma 2/GABRG2 antibody [EPR25325-5] (AB288564)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Neuro-2a cells labelling GABA A Receptor gamma 2/GABRG2 with ab288564 at 1/500 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in Neuro-2a 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 ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-GABA A Receptor gamma 2/GABRG2 antibody [EPR25325-5] (AB288564)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SH-SY5Y cells labelling GABA A Receptor gamma 2/GABRG2 with ab288564 at 1/500 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in SH-SY5Y cell line.Negative control : 293T (PMID : 27864268） 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 ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution.

• IP

Lab

Immunoprecipitation - Anti-GABA A Receptor gamma 2/GABRG2 antibody [EPR25325-5] (AB288564)

GABA A Receptor gamma 2/GABRG2 was immunoprecipitated from 0.35 mg Mouse brain tissue lysate 10ug with ab288564 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab288564 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse brain tissue lysate 10ug

Lane 2 : ab288564 IP in Mouse brain tissue lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab288564 in Mouse brain tissue lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 101 seconds

Observed MW(KDa) 50

All lanes:

Immunoprecipitation - Anti-GABA A Receptor gamma 2/GABRG2 antibody [EPR25325-5] (ab288564)

Predicted band size: 54 kDa

Observed band size: 50 kDa

false

• WB

Lab

Western blot - Anti-GABA A Receptor gamma 2/GABRG2 antibody [EPR25325-5] (AB288564)

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

The blot of Lane 3&5 was developed using a higher sensitivity ECL substrate.

Exposure time : Lane 1, 2 &4 : 147 seconds Lane 3 & 5 : 3 minutes

All lanes:

Western blot - Anti-GABA A Receptor gamma 2/GABRG2 antibody [EPR25325-5] (ab288564) at 1/1000 dilution

Lane 1:

Rat brain tissue lysate at 20 µg

Lane 2:

Rat cerebral cortex tissue lysate at 20 µg

Lane 3:

Mouse brain tissue lysate at 20 µg

Lane 4:

Mouse cerebellum tissue lysate at 20 µg

Lane 5:

Neuro-2a (mouse neuroblastoma neuroblast), whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 54 kDa

Observed band size: 54 kDa

false

• WB

Lab

Western blot - Anti-GABA A Receptor gamma 2/GABRG2 antibody [EPR25325-5] (AB288564)

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

Negative control : 293T (PMID : 27864268).

Exposure time : 147 seconds

All lanes:

Western blot - Anti-GABA A Receptor gamma 2/GABRG2 antibody [EPR25325-5] (ab288564) at 1/1000 dilution

Lane 1:

SH-SY5Y (human neuroblastoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

HEK-293T (Human embryonic kidney epithelial cell), whole cell lysate at 20 µg

Lane 3:

A-172 (human brain glioblastoma), whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 54 kDa

Observed band size: 54 kDa

false