Anti-FOXK1/MNF antibody

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-FOXK1/MNF antibody (AB18196)

ICC/IF image of ab18196 stained Hek293 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab18196, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

• WB

AbReview36868****

Western blot - Anti-FOXK1/MNF antibody (AB18196)

All lanes:

Western blot - Anti-FOXK1/MNF antibody (ab18196) at 1/5000 dilution

Lane 1:

Human H1299 cell lysate at 20 µg

Lane 2:

Mouse embryonic stem cell lysate at 20 µg

Secondary

All lanes:

HRP-conjugate Goat anti-rabbit IgG polyclonal at 1/5000 dilution

Predicted band size: 75 kDa

Observed band size: 100 kDa

true

Exposure time: 1min

This image is courtesy of an anonymous Abreview

• IP

Lab

Immunoprecipitation - Anti-FOXK1/MNF antibody (AB18196)

FOXK1/MNF - ChIP Grade was immunoprecipitated using 0.5mg Hela whole cell extract, 5μg of Rabbit polyclonal to FOXK1/MNF and 50μl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40μl SDS loading buffer and incubated for 10min at 70^oC; 10μl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab18196.
Secondary : Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band : 96kDa : FOXK1 / MNF.

All lanes:

Immunoprecipitation - Anti-FOXK1/MNF antibody (ab18196)

Predicted band size: 75 kDa

false

• WB

Project

Western blot - Anti-FOXK1/MNF antibody (AB18196)

All lanes:

Western blot - Anti-FOXK1/MNF antibody (ab18196) at 1/250 dilution

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 2:

HeLa (Human epithelial carcinoma cell line) Nuclear Lysate at 10 µg

Lane 3:

HepG2 (Human hepatocellular liver carcinoma cell line) Nuclear Lysate at 10 µg

Lane 4:

Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 10 µg

Lane 5:

Jurkat nuclear extract lysate (<a href='/en-us/products/unavailable/jurkat-nuclear-extract-lysate-ab14844'>ab14844</a>) at 10 µg

Lane 6:

K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell Lysate at 10 µg

Lane 7:

Western blot - SW480 whole cell lysate (<a href='/en-us/products/cell-lysates/sw480-whole-cell-lysate-ab3957'>ab3957</a>) at 10 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Predicted band size: 75 kDa

Observed band size: 95 kDa

true

Exposure time: 5min