Anti-Ikaros antibody [EPR13790]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Ikaros antibody [EPR13790] (AB191394)

Intracellular flow cytometric analysis of MOLT4 cells (paraformaldehyde-fixed, 2%)labeling Ikaros with ab191394 at 1/130 dilution (red) or a rabbit IgG (negative) (green), followed by Goat anti rabbit IgG (FITC) secondary at 1/150 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ikaros antibody [EPR13790] (AB191394)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Ikaros with ab191394 at 1/1400 dilution followed by pre-diluted HRP Polymer for Rabbit IgG secondary antibody and counter-stained with Hematoxylin.
Inset : Negative control : using PBS instead of primary antibody.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ikaros antibody [EPR13790] (AB191394)

Immunohistochemical analysis of paraffin-embedded Human thymus tissue labeling Ikaros with ab191394 at 1/1400 dilution followed by pre-diluted HRP Polymer for Rabbit IgG secondary antibody and counter-stained with Hematoxylin.
Inset : Negative control : using PBS instead of primary antibody.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

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Western blot - Anti-Ikaros antibody [EPR13790] (AB191394)

Based on the sequence analysis, ab191394 recognizes seven isoforms with the predicted MWs of 58KDa, 48KDa, 48KDa, 43KDa, 41KDa, 32KDa and 53KDa, respectively.

All lanes:

Western blot - Anti-Ikaros antibody [EPR13790] (ab191394) at 1/10000 dilution

Lane 1:

Raji cell lysate at 20 µg

Lane 2:

Ramos cell lysate at 20 µg

Lane 3:

MOLT4 cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 57 kDa

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• WB

Lab

Western blot - Anti-Ikaros antibody [EPR13790] (AB191394)

Lanes 1 - 4 : Merged signal (red and green). Green - ab191394 observed at 50-70 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab191394 was shown to react with Ikaros in wild-type Jurkat cells in western blot with loss of signal observed in IKZF1 knockout sample. Wild-type and IKZF1 knockout Jurkat cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab191394 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Ikaros antibody [EPR13790] (ab191394) at 1/10000 dilution

Lane 1:

Wild-type Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

Lane 2:

IKZF1 knockout Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

Lane 2:

Western blot - Human IKZF1 knockout Jurkat cell line (<a href='/en-us/products/cell-lines/human-ikzf1-knockout-jurkat-cell-line-ab274920'>ab274920</a>)

Lane 3:

Daudi (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg

Lane 4:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Predicted band size: 57 kDa

Observed band size: 50-70 kDa

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