Anti-GCLC antibody [EPR20078]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-GCLC antibody [EPR20078] (AB207777)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling GCLC with ab207777 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and weak nuclear staining on HepG2 cell line. The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-GCLC antibody [EPR20078] (AB207777)

GCLC was immunoprecipitated from 0.35 mg of HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate with ab207777 at 1/40 dilution.

Western blot was performed from the immunoprecipitate using ab207777 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1 : HepG2 whole cell lysate 10μg (Input).

Lane 2 : ab207777 IP in HepG2 whole cell lysate.

Lane 3 : Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab207777 in HepG2 whole cell lysate.

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

Exposure time : 30 seconds.

All lanes:

Immunoprecipitation - Anti-GCLC antibody [EPR20078] (ab207777)

Predicted band size: 73 kDa

false

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-GCLC antibody [EPR20078] (AB207777)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed PC-12 (Rat adrenal gland pheochromocytoma cell line) cells labeling GCLCwith ab207777 at 1/60 dilution (red) compared with aRabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat Anti-Rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-GCLC antibody [EPR20078] (AB207777)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized PC-12 (Rat adrenal gland pheochromocytoma cell line) cells labeling GCLC with ab207777 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and weak nuclear staining on PC-12 cell line. The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.

• WB

Supplier Data

Western blot - Anti-GCLC antibody [EPR20078] (AB207777)

Blocking/Dilution buffer : 5% NFDM/TBST.

Lanes 1 - 2:

Western blot - Anti-GCLC antibody [EPR20078] (ab207777) at 1/2000 dilution

Lanes 3 - 5:

Western blot - Anti-GCLC antibody [EPR20078] (ab207777) at 1/1000 dilution

Lane 1:

HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 10 µg

Lane 2:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 10 µg

Lane 3:

A549 (Human lung carcinoma cell line) whole cell lysate at 10 µg

Lane 4:

Human lung lysate at 10 µg

Lane 5:

Human fetal liver lysate at 10 µg

Secondary

Lanes 1 - 3:

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

Lanes 4 - 5:

Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 73 kDa

Observed band size: 73 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-GCLC antibody [EPR20078] (AB207777)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1-3 : 3 minutes; Lane 4 : 15 seconds; Lane 5 : 3 seconds; Lane 6 : 5 seconds.

Lanes 1 - 2:

Western blot - Anti-GCLC antibody [EPR20078] (ab207777) at 1/2000 dilution

Lanes 3 - 6:

Western blot - Anti-GCLC antibody [EPR20078] (ab207777) at 1/1000 dilution

Lane 1:

C6 (Rat glial tumor cell line) whole cell lysate at 10 µg

Lane 2:

PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg

Lane 3:

RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

Lane 4:

Rat liver lysate at 10 µg

Lane 5:

Mouse liver lysate at 10 µg

Lane 6:

Mouse spleen lysate at 10 µg

Secondary

All lanes:

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

Predicted band size: 73 kDa

Observed band size: 73 kDa

false