Anti-N myc interactor/NMI antibody [EPR11065(2)] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N myc interactor/NMI antibody [EPR11065(2)] - BSA and Azide free (AB250698)

This data was developed using ab183724, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling N myc interactor/NMI with ab183724 at 1/50 dilution. The slide is counterstained with Hematoxylin.

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-N myc interactor/NMI antibody [EPR11065(2)] - BSA and Azide free (AB250698)

This data was developed using ab183724, the same antibody clone in a different buffer formulation.

Immunofluorescence analysis of acetone-fixed HeLa cells labeling N myc interactor/NMI with ab183724 at 1/100 dilution. Goat anti-rabbit IgG (Alexa Fluor® 488) at 1/200 dilution was used as the secondary antibody (green). The slide on the right is stained with Dapi (blue).

• IP

Supplier Data

Immunoprecipitation - Anti-N myc interactor/NMI antibody [EPR11065(2)] - BSA and Azide free (AB250698)

This data was developed using ab183724, the same antibody clone in a different buffer formulation.

Western blot analysis of K562 cell lysate precipitated with ab183724 at 1/50 dilution. A Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated secondary antibody at 1/1000 dilution was then used. The blocking buffer and dilution buffer was 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-N myc interactor/NMI antibody [EPR11065(2)] (<a href='/en-us/products/primary-antibodies/n-myc-interactor-nmi-antibody-epr110652-ab183724'>ab183724</a>)

Predicted band size: 35 kDa

false

• WB

Lab

Western blot - Anti-N myc interactor/NMI antibody [EPR11065(2)] - BSA and Azide free (AB250698)

This data was developed using ab183724, the same antibody clone in a different buffer formulation.

Lane 1 : Wild-type HAP1 cell lysate (20 μg)

Lane 2 : N myc interactor/NMI knockout HAP1 cell lysate (20 μg)

Lane 3 : HeLa cell lysate (20 μg)

Lane 4 : K562 cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab183724 observed at 39 kDa. Red - loading control, ab18058, observed at 124 kDa.

ab183724 was shown to specifically react with N myc interactor/NMI when N myc interactor/NMI knockout samples were used. Wild-type and N myc interactor/NMI knockout samples were subjected to SDS-PAGE. ab183724 and ab18058 (loading control to Vinculin) were diluted at 1/2000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-N myc interactor/NMI antibody [EPR11065(2)] (<a href='/en-us/products/primary-antibodies/n-myc-interactor-nmi-antibody-epr110652-ab183724'>ab183724</a>)

Predicted band size: 35 kDa

false

• WB

Supplier Data

Western blot - Anti-N myc interactor/NMI antibody [EPR11065(2)] - BSA and Azide free (AB250698)

This data was developed using ab183724, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-N myc interactor/NMI antibody [EPR11065(2)] (<a href='/en-us/products/primary-antibodies/n-myc-interactor-nmi-antibody-epr110652-ab183724'>ab183724</a>) at 1/20000 dilution

Lane 1:

K562 cell lysate at 20 µg

Lane 2:

HeLa cell lysate at 20 µg

Lane 3:

HepG2 cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 35 kDa

false

• WB

Lab

Western blot - Anti-N myc interactor/NMI antibody [EPR11065(2)] - BSA and Azide free (AB250698)

This data was developed using ab183724, the same antibody clone in a different buffer formulation.

Lanes 1-4 : Merged signal (red and green). Green - ab183724 observed at 39 kDa. Red - loading control ab8245 observed at 36 kDa.

ab183724 Anti-N myc interactor/NMI antibody [EPR11065(2)] was shown to specifically react with N myc interactor/NMI in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267013 (knockout cell lysate ab258077) was used. Wild-type and N myc interactor/NMI knockout samples were subjected to SDS-PAGE. ab183724 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-N myc interactor/NMI antibody [EPR11065(2)] (<a href='/en-us/products/primary-antibodies/n-myc-interactor-nmi-antibody-epr110652-ab183724'>ab183724</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

NMI knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human NMI (N myc interactor/NMI) knockout A549 cell line (<a href='/en-us/products/cell-lines/human-nmi-n-myc-interactor-nmi-knockout-a549-cell-line-ab267013'>ab267013</a>)

Lane 3:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 4:

K562 (Human chronic myelogenous leukemia lymphoblast cell line ) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 35 kDa

Observed band size: 39 kDa

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