Anti-Nkx2.5 antibody [EPR20168] - BSA and Azide free

• WB

Lab

Western blot - Anti-Nkx2.5 antibody [EPR20168] - BSA and Azide free (AB228335)

False colour image of Western blot : Anti-Nkx2.5 antibody [EPR20168] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab205263 was shown to bind specifically to Nkx2.5. A band was observed at 40, 30, 25 kDa in wild-type HEK-293 cell lysates with no signal observed at this size in NKX2-5 knockout cell line ab261899 (knockout cell lysate ab261716). Bands at 37 and 35 kDa are non-specific believed to be NKX2-2 and NKX2-6. To generate this image, wild-type and NKX2-5 knockout HEK-293 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-Nkx2.5 antibody [EPR20168] (<a href='/en-us/products/primary-antibodies/nkx25-antibody-epr20168-ab205263'>ab205263</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293 cell lysate at 20 µg

Lane 2:

NKX2-5 knockout HEK-293 cell lysate at 20 µg

Lane 2:

Western blot - Human NKX2-5 knockout HEK-293 cell line (<a href='/en-us/products/cell-lines/human-nkx2-5-knockout-hek-293-cell-line-ab261899'>ab261899</a>)

Lane 3:

HeLa cell lysate at 20 µg

Predicted band size: 35 kDa

Observed band size: 25 kDa,30 kDa,40 kDa

false

• WB

Supplier Data

Western blot - Anti-Nkx2.5 antibody [EPR20168] - BSA and Azide free (AB228335)

Blocking/Dilution : 5% NFDM/TBST

The molecular masses observed are consistent with what has been described in the literature (PMID : 21447557)

Negative control : MCF7 (PMID : 15340050).

The blot was developed on a BIO-RAD^® ChemiDoc™ MP instrument.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab205263).

All lanes:

Western blot - Anti-Nkx2.5 antibody [EPR20168] (<a href='/en-us/products/primary-antibodies/nkx25-antibody-epr20168-ab205263'>ab205263</a>) at 1/1000 dilution

Lane 1:

Human fetal heart muscle tissue lysate at 20 µg

Lane 2:

HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

MCF7 (human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/2000 dilution

Predicted band size: 35 kDa

Observed band size: 30-42 kDa

true

Exposure time: 3min