Anti-ILF3 antibody [EPR3626]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-ILF3 antibody [EPR3626] (AB92355)

Overlay histogram showing HeLa cells stained with unpurified ab92355 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab92355, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monclonal) (1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-ILF3 antibody [EPR3626] (AB92355)

Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling ILF3 with purified ab92355 at 1/60 dilution (10μg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor^® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-ILF3 antibody [EPR3626] (AB92355)

Immunofluorescence staining of HeLa cells with purified ab92355 at a working dilution of 1/500, counter-stained with DAPI. The secondary antibody was an Alexa Fluor^® 488 conjugated goat anti-rabbit (ab150077), used at a dilution of 1/1000. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative control is shown in bottom right hand panel - for the negative control, PBS was used instead of the primary antibody.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ILF3 antibody [EPR3626] (AB92355)

ab92355 (unpurified) at 1/100 dilution staining ILF3 in paraffin-embedded Human kidney tissue, by immunohistochemistry.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• WB

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Western blot - Anti-ILF3 antibody [EPR3626] (AB92355)

Blocking and diluting buffer : 5% NFDM/TBST.
The two bands are reported by PMID : 22842455 and 27872311.

All lanes:

Western blot - Anti-ILF3 antibody [EPR3626] (ab92355) at 1/100000 dilution

Lane 1:

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

Lane 2:

SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysates at 20 µg

Lane 3:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 20 µg

Lane 4:

Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysates at 20 µg

Lane 5:

C6 (Rat glial tumor glial cell) whole cell lysates 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: 95 kDa

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• WB

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Western blot - Anti-ILF3 antibody [EPR3626] (AB92355)

All lanes:

Western blot - Anti-ILF3 antibody [EPR3626] (ab92355) at 1/100000 dilution

Lane 1:

Raji cell lysate at 10 µg

Lane 2:

HeLa cell lysate at 10 µg

Lane 3:

K562 cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit antibody at 1/2000 dilution

Predicted band size: 95 kDa

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