Anti-Oct-2 antibody [EPR12482]

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Oct-2 antibody [EPR12482] (AB179808)

Immunocytochemistry/ Immunofluorescence analysis of Ramos (Human Burkitt's lymphoma B lymphocyte) cells labeling Oct-2 with purified ab179808 at 1/200 (2.7 μg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 μg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 μg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Oct-2 antibody [EPR12482] (AB179808)

Intracellular Flow Cytometry analysis of Raji (Human Burkitt's lymphoma B lymphocyte) cells labeling Oct-2 with purified ab179808 at 1/50 dilution (10μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• IP

Unknown

Immunoprecipitation - Anti-Oct-2 antibody [EPR12482] (AB179808)

Purified ab179808 at 1/30 dilution (2ug) immunoprecipitating Oct-2 in Raji whole cell lysate.
Lane 1 (input) : Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate (10μg)
Lane 2 (+) : ab179808 + Raji whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab179808 in Raji whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000) was used for Western blotting.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : 58 kDa

All lanes:

Immunoprecipitation - Anti-Oct-2 antibody [EPR12482] (ab179808)

Predicted band size: 51 kDa

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• WB

Lab

Western blot - Anti-Oct-2 antibody [EPR12482] (AB179808)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/1000,000 dilution.

Negative samples : HeLa, A549, SW480 (PMID : 20473321).

All lanes:

Western blot - Anti-Oct-2 antibody [EPR12482] (ab179808) at 1/1000 dilution

Lane 1:

Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 2:

Ramos (human Burkitt's lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 3:

Daudi (human Burkitt's lymphoma lymphoblast) whole cell lysate at 20 µg

Lane 4:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

SW480 (human colorectal 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 1/20000 dilution

Observed band size: 65 kDa,36 kDa

false

Exposure time: 10s

• WB

Unknown

Western blot - Anti-Oct-2 antibody [EPR12482] (AB179808)

Blocking/Diluting buffer : 5% NFDM/TBST

The observed bands are consistent with what are described in PMID : 26993806.

All lanes:

Western blot - Anti-Oct-2 antibody [EPR12482] (ab179808) at 1/5000 dilution

Lane 1:

Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 2:

Mouse brain lysate at 20 µg

Lane 3:

Rat brain 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 1/20000 dilution

Predicted band size: 51 kDa

Observed band size: 65 kDa

false