Anti-eIF4EBP1 (phospho T46) antibody [19HCLC]

• ICC

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Immunocytochemistry - Anti-eIF4EBP1 (phospho T46) antibody [19HCLC] (AB277767)

Immunofluorescent analysis of 4EBP1 (pT46) was done on 70% confluent log phase HeLa (Human epithelial cell line from cervix adenocarcinoma) cells. The cells were fixed with 4% paraformaldehyde for 15 minutes; permeabilized with 0.25% Triton X-100 for 10 minutes followed by blocking with 5% BSA for 1 hour at room temperature. The cells were incubated with ab277767 at a dilution of 2 μg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor^® 488 Goat anti-Rabbit IgG Secondary Antibody at a dilution of 1/400 for 30 minutes at room temperature (Panel a : green). Nuclei (Panel b : blue) were stained with with DAPI. F-actin (Panel c : red) was stained with Alexa Fluor^® 594 Phalloidin. Panel d is a merged image showing nuclear localization of 4EBP1 (pT46). Panel e shows competition with 4EBP1 (pT46) peptide. The images were captured at 20X magnification.

• ICC

Supplier Data

Immunocytochemistry - Anti-eIF4EBP1 (phospho T46) antibody [19HCLC] (AB277767)

Immunofluorescent analysis of Phospho-4EBP1 pThr46 in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells using ab277767 followed by detection using an Alexa Fluor^® 488-conjugated Goat anti-Rabbit secondary antibody (green) (Image A). Nuclei were stained using DAPI (Image B) and actin stained with Alexa Fluor^® 594 phalloidin (red) (image C). Image D is a composite image showing nuclear localization of phosphorylated 4EBP1 and Image E is a composite image of cells showing inhibition of antibody binding after competition with the phosphorylated peptide.

• WB

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Western blot - Anti-eIF4EBP1 (phospho T46) antibody [19HCLC] (AB277767)

All lanes:

Western blot - Anti-eIF4EBP1 (phospho T46) antibody [19HCLC] (ab277767) at 2 µg/mL

Lane 1:

HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 30 µg

Lane 2:

HEK-293 whole cell lysate treated for 10 minutes with 100 nM/mL of EGF at 30 µg

Lane 3:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 30 µg

Lane 4:

MCF7 whole cell lysate treated for 15 minutes with 150 µg/mL of Insulin at 30 µg

Secondary

All lanes:

Goat anti-Rabbit IgG - HRP at 1/5000 dilution

Predicted band size: 13 kDa

false

• WB

Supplier Data

Western blot - Anti-eIF4EBP1 (phospho T46) antibody [19HCLC] (AB277767)

All lanes:

Western blot - Anti-eIF4EBP1 (phospho T46) antibody [19HCLC] (ab277767) at 2 µg/mL

Lane 1:

HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysatetreated with EGF

Lane 2:

Preincubation with the phosphopeptide, HEK-293 whole cell lysate

Lane 3:

NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate treated with PDGF

Lane 4:

Preincubation with the phosphopeptide, NIH/3T3 whole cell lysate

Predicted band size: 13 kDa

false