Anti-TIM 4 antibody [EPR22304-3]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-TIM 4 antibody [EPR22304-3] (AB222093)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized TIM 4 transfected HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) cells labeling TIM 4 (Green) with ab222093 at 1/100 dilution, followed by AlexaFluor^®488 Goat anti-Rabbit secondary (ab150077) antibody at 1/1000 dilution. Confocal image showing positive staining in HEK-293T cells transfected with a myc-tagged TIM 4 expression construct. myc-Tag (9B11) Mouse mAb (Alexa Fluor^® 594 Conjugate) (ab223894) was used as the counterstain (Red) at 1/100 dilution. The nuclear counterstain was DAPI (Blue).

Negative control : Myc only transfected HEK-293T cells.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is AlexaFluor^®488 Goat anti-Rabbit secondary (ab150077).

• Flow Cyt

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Flow Cytometry - Anti-TIM 4 antibody [EPR22304-3] (AB222093)

Flow cytometric analysis of 4% paraformaldehyde-fixed HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with a myc-tagged TIM 4 expression construct labeling TIM 4 with ab222093 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730, Black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody, Blue).

Cells were surface stained with ab222093. Then fixed with 2% PFA for 10min, followed by intracellular staining of anti-myc tag conjugated to Alexa Fluor^®647.

Gated on myc -positive population.

• WB

Supplier Data

Western blot - Anti-TIM 4 antibody [EPR22304-3] (AB222093)

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

In Western blot, Anti-GFP antibody [E385] (ab32146) staining at 1/10000 dilution.

In Western blot, Anti-TIM 4 antibody [EPR22304-3] (ab222093) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-HA tag antibody [RM1058] (<a href='/ko/products/primary-antibodies/ha-tag-antibody-rm1058-ab314237'>ab314237</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney) transfected with an empty vector (vector control), containing a myc-GFP-tag, whole cell lysate at 20 µg

Lane 2:

293T transfected with TIM 4 (WT) expression vector containing a HA-myc-His-GFP-tag and PDGFR TM domain, whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 78 kDa,36 kDa

false

Exposure time: 37s