Anti-NM23A antibody [EPR3036] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NM23A antibody [EPR3036] - BSA and Azide free (AB247543)

This data was developed using ab92327, the same antibody clone in a different buffer formulation.

ab92327, at a dilution of 1/100, staining NM23A in formalin fixed, paraffin embedded Human lung adenocarcinoma tissue by Immunohistochemistry. Detection : DAB staining

Heat mediated antigen retrieval was performed via the pressure cooker method before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-NM23A antibody [EPR3036] - BSA and Azide free (AB247543)

This data was developed using ab92327, the same antibody clone in a different buffer formulation.

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling NM23A with unpurified ab92327 at 1/50 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti-rabbit IgG (Alexa Fluor^® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

• WB

Unknown

Western blot - Anti-NM23A antibody [EPR3036] - BSA and Azide free (AB247543)

This data was developed using ab92327, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-NM23A antibody [EPR3036] (<a href='/en-us/products/primary-antibodies/nm23a-antibody-epr3036-ab92327'>ab92327</a>) at 1/5000 dilution

Lane 1:

MCF-7 cell lysate at 10 µg

Lane 2:

A375 cell lysate at 10 µg

Lane 3:

HeLa cell lysate at 10 µg

Lane 4:

Jurkat cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 17 kDa

false

• WB

Lab

Western blot - Anti-NM23A antibody [EPR3036] - BSA and Azide free (AB247543)

This data was developed using the same antibody clone in a different buffer formulation (ab92327).

Lanes 1-4 : Merged signal (red and green). Green - ab92327 observed at 17 kDa. Red - loading control ab8245 observed at 36 kDa.

ab92327 Anti-NM23A antibody [EPR3036] was shown to specifically react with NM23A in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266215 (knockout cell lysate ab258075) was used. Wild-type and NM23A knockout samples were subjected to SDS-PAGE. ab92327 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-NM23A antibody [EPR3036] (<a href='/en-us/products/primary-antibodies/nm23a-antibody-epr3036-ab92327'>ab92327</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

NME1 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human NME1 (NM23A) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-nme1-nm23a-knockout-hek-293t-cell-line-ab266215'>ab266215</a>)

Lane 3:

MCF7 cell lysate at 20 µg

Lane 4:

Jurkat 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: 17 kDa

Observed band size: 17 kDa

false