Anti-CLIC1 antibody [EPR22907-50]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CLIC1 antibody [EPR22907-50] (AB229917)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized C2C12 (mouse myoblasts myoblast) cells labelling CLIC1 with at 1/100 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic and membranous staining in C2C12 cell line. ab195889 Anti-alpha Tubulin antibody [DM1A] - 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 : Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution.

• WB

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Western blot - Anti-CLIC1 antibody [EPR22907-50] (AB229917)

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

Exposure times.
Lane 1 : 26 seconds;
Lane 2 : 10 seconds;
Lanes 3-4 : 3 minutes.

All lanes:

Western blot - Anti-CLIC1 antibody [EPR22907-50] (ab229917) at 1/1000 dilution

Lane 1:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 2:

C2C12 (mouse myoblasts myoblast) whole cell lysate at 20 µg

Lane 3:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

MDA-MB-231 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 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: 26 kDa

Observed band size: 27 kDa

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• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-CLIC1 antibody [EPR22907-50] (AB229917)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized K562 (human chronic myelogenous leukemia lymphoblast) cells labelling CLIC1 with at 1/100 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in K-562 cell line. ab195889 Anti-alpha Tubulin antibody [DM1A] - 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 : Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-CLIC1 antibody [EPR22907-50] (AB229917)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized K-562 (Human chronic myelogenous leukemia lymphoblast) cells labelling CLIC1 with ab229917 at 1/500 dilution (0.1�g) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^� 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-CLIC1 antibody [EPR22907-50] (AB229917)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized C2C12 (Mouse myoblasts myoblast) cells labelling CLIC1 with ab229917 at 1/500 dilution (0.1�g) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^� 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• IP

Unknown

Immunoprecipitation - Anti-CLIC1 antibody [EPR22907-50] (AB229917)

CLIC1 was immunoprecipitated from 0.35mg Mouse placenta lysate with ab229917 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab229917 at 1/1000 dilution (0.5 μg/ml). VeriBlot for IP Detection Reagent (HRP) (ab131366) was used as the secondary antibody at 1/5000 dilution.

Lane 1 : Mouse placenta lysate 10μg

Lane 2 : ab229917 IP in Mouse placenta lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab229917 in Mouse placenta lysate.

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

Exposure time : 30 seconds.

All lanes:

Immunoprecipitation - Anti-CLIC1 antibody [EPR22907-50] (ab229917)

Predicted band size: 26 kDa

Observed band size: 27 kDa

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• WB

Unknown

Western blot - Anti-CLIC1 antibody [EPR22907-50] (AB229917)

Exposure times.
Lanes 1-2 : 15 seconds;
Lane 3 : 3 minutes;
Lanes 4-5 : 48 seconds.

Blocking buffer and concentration : 5% NFDM/TBST.

All lanes:

Western blot - Anti-CLIC1 antibody [EPR22907-50] (ab229917) at 1/1000 dilution

Lane 1:

Mouse kidney lysate at 10 µg

Lane 2:

Mouse spleen lysate at 10 µg

Lane 3:

Rat spleen lysate at 10 µg

Lane 4:

C6 (rat glial tumor glial cell) whole cell lysate at 10 µg

Lane 5:

RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell 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: 26 kDa

Observed band size: 27 kDa

false

• WB

Unknown

Western blot - Anti-CLIC1 antibody [EPR22907-50] (AB229917)

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-CLIC1 antibody [EPR22907-50] (ab229917) at 1/1000 dilution

Lane 1:

Human fetal heart lysate at 20 µg

Lane 2:

Human fetal kidney lysate at 20 µg

Lane 3:

Human fetal spleen lysate at 20 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 26 kDa

Observed band size: 27 kDa

false

Exposure time: 15s