Anti-IL-7 antibody [EPR27289-137]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-7 antibody [EPR27289-137] (AB324645)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human epithelial cell line from embryonic kidney) transfected with a human IL7 expression vector containing a Myc-His tag, (B) HEK-293T transfected with a mouse IL7 expression vector containing a Myc-His tag and (C) HEK-293T transfected with empty vector containing a Myc-His tag labeling IL-7 with ab324645 at 1/500 (1.014 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) HEK-293T (human epithelial cell line from embryonic kidney) transfected with a human IL7 expression vector containing a Myc-His tag, (B) HEK-293T transfected with a mouse IL7 expression vector containing a Myc-His tag, and no staining on (C) HEK-293T transfected with empty vector containing a Myc-His tag. The section was incubated with ab324645 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-IL-7 antibody [EPR27289-137] (AB324645)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling IL-7 with ab324645 at 1/1000 (0.507 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing positive staining in 293T (shown in green) cells transfected with a mouse Interleukin-7 or a human Interleukin-7 expression vector containing a myc-His-tag®. The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab223894 Anti-Myc tag mouse monoclonal antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/100 (5 ug/ml) dilution (Magenta).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-IL-7 antibody [EPR27289-137] (AB324645)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype (Left) / 293T cells transfected with a human IL-7 expression vector containing a myc-His-tag® (Middle) / 293T cells transfected with an empty expression vector containing a myc-His-tag® (Right) cells labelling IL-7 with ab324645 at 1/500 dilution (0.1ug) / Middle and Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control.

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Cells are co-stained with Myc tag conjugated to Alexa Fluor®647.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-IL-7 antibody [EPR27289-137] (AB324645)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype (Left) / 293T cells transfected with a mouse IL-7 expression vector containing a myc-His-tag® (Middle) / 293T cells transfected with an empty expression vector containing a myc-His-tag® (Right) cells labelling IL-7 with ab324645 at 1/500 dilution (0.1ug) / Middle and Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control.

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Cells are co-stained with Myc tag conjugated to Alexa Fluor®647.

• WB

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Western blot - Anti-IL-7 antibody [EPR27289-137] (AB324645)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lane 1, 2 : 26 seconds Lane 3, 4 : 180 seconds

All lanes:

Western blot - Anti-IL-7 antibody [EPR27289-137] (ab324645) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a Myc-his tag, whole cell lysate at 7 µg

Lane 2:

293T cells transfected with a human IL-7 expression vector containing a Myc-his tag, whole cell lysate at 7 µg

Lane 3:

293T cells transfected with an empty vector containing a Myc-his tag, whole cell lysate at 4 µg

Lane 4:

293T cells transfected with a mouse IL-7 expression vector containing a Myc-his tag, whole cell lysate at 4 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

Observed band size: 20 kDa,36 kDa,20-25 kDa

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