Anti-FOXO4/AFX antibody [EPR5442]

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-FOXO4/AFX antibody [EPR5442] (AB128908)

Intracellular flow cytometric analysis of FOXO4/AFX in permeabilized HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) cells using ab128908 at 1/100 dilution (Red) compared to a non-specific negative control antibody (green).

• IP

Unknown

Immunoprecipitation - Anti-FOXO4/AFX antibody [EPR5442] (AB128908)

Purified ab128908 at 1/100 dilution (2μg) immunoprecipitating FOXO4/AFX in 293T whole cell lysate.
Lane 1 (input) : 293T (Human embryonic kidney epithelial cell) whole cell lysate 10μg
Lane 2 (+) : ab128908 + 293T whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab128908 in 293T whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : 65 kDa

All lanes:

Immunoprecipitation - Anti-FOXO4/AFX antibody [EPR5442] (ab128908)

Predicted band size: 53 kDa

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• WB

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Western blot - Anti-FOXO4/AFX antibody [EPR5442] (AB128908)

All lanes:

Western blot - Anti-FOXO4/AFX antibody [EPR5442] (ab128908) at 1/1000 dilution

Lane 1:

293T lysate at 10 µg

Lane 2:

Fetal heart lysate at 10 µg

Lane 3:

Fetal muscle lysate at 10 µg

Lane 4:

Fetal lung lysate at 10 µg

Lane 5:

HuT-78 lysate at 10 µg

Secondary

All lanes:

Goat-anti-rabbit HRP at 1/2000 dilution

Predicted band size: 53 kDa

Observed band size: 65 kDa

false

• WB

Supplier Data

Western blot - Anti-FOXO4/AFX antibody [EPR5442] (AB128908)

Blocking and diluting buffer and concentration : 5% NFDM/TBST The bands above 100 kDa are unknown. In Western blot, anti-FOXO4 antibody (ab128908) staining at 1/1000 dilution. Exposure time : 59 seconds.

All lanes:

Western blot - Anti-FOXO1A (pT24)+FOXO3A (pT32)+FOXO4 (pT28)+FOXO6 (pT26) antibody [EPR28359-78] (<a href='/en-us/products/primary-antibodies/foxo1a-pt24foxo3a-pt32foxo4-pt28foxo6-pt26-antibody-epr28359-78-ab312326'>ab312326</a>) at 1/1000 dilution

Lane 1:

Jurkat (human t cell leukemia t lymphocyte from peripheral blood) starved overnight, then treated with 50 uM LY294092 for 30min whole cell lysate (untreated membrane) at 20 µg

Lane 2:

Jurkat starved overnight, then treated with 20% serum and 100nM Calycin A for 30min whole cell lysate (untreated membrane) at 20 µg

Lane 3:

Jurkat starved overnight, then treated with 20% serum and 100nM Calycin A for 30min whole cell lysate (phosphatase treated membrane) 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

Observed band size: 75-80 kDa

false

Exposure time: 59s

• WB

Unknown

Western blot - Anti-FOXO4/AFX antibody [EPR5442] (AB128908)

All lanes:

Western blot - Anti-FOXO4/AFX antibody [EPR5442] (ab128908) at 1/1000 dilution

Lane 1:

293T transfected with FOXO4/AFX

Lane 2:

293T cell lysate

Secondary

All lanes:

Goat-anti-rabbit HRP at 1/2000 dilution

Predicted band size: 53 kDa

Observed band size: 63 kDa

false

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-FOXO4/AFX antibody [EPR5442] (AB128908)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.