Anti-Connexin 45/GJA7/Cx45 antibody [EPR26705-152]

• IP

Supplier Data

Immunoprecipitation - Anti-Connexin 45/GJA7/Cx45 antibody [EPR26705-152] (AB316742)

Connexin 45/GJA7/Cx45 was immunoprecipitated from 0.35 mg IMR-32 (human neuroblastoma neuroblast) whole cell lysate with ab316742 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab316742 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : IMR-32 (human neuroblastoma neuroblast) whole cell lysate
Lane 2 : ab316742 IP in IMR-32 (human neuroblastoma neuroblast) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab316742 in IMR-32 whole cell lysate

All lanes:

Immunoprecipitation - Anti-Connexin 45/GJA7/Cx45 antibody [EPR26705-152] (ab316742) at 1/30 dilution

All lanes:

IMR-32 (human neuroblastoma neuroblast) whole cell lysate with 5% NFDM/TBST

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 8s

• WB

Supplier Data

Western blot - Anti-Connexin 45/GJA7/Cx45 antibody [EPR26705-152] (AB316742)

Negative control : MCF7, HT-29.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lane 1 : 180 seconds； Lanes 2-5 : 37 seconds.

All lanes:

Western blot - Anti-Connexin 45/GJA7/Cx45 antibody [EPR26705-152] (ab316742) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 2:

NCI-H1299 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 3:

IMR-32 (human neuroblastoma neuroblast) whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 4:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

Lane 5:

HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg with 5% NFDM/TBST

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: 45 kDa,36 kDa

false

• WB

Lab

Western blot - Anti-Connexin 45/GJA7/Cx45 antibody [EPR26705-152] (AB316742)

Western blot : Anti-Connexin 45/GJA7/Cx45 antibody [EPR26705-152] ab316742 staining at 1/1000 dilution, shown in green; Mouse anti-CANX (ab238078) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 45 kDa in Wild-type A549 UNBOILED cell lysates with no signal observed at this size in GJC1 knockout A549 UNBOILED cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc 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 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-Connexin 45/GJA7/Cx45 antibody [EPR26705-152] (ab316742) at 1/1000 dilution

Lane 1:

Wild-type A549 UNBOILED at 20 µg

Lane 2:

GJC1 knockout A549 UNBOILED at 20 µg

Lane 3:

U-2 OS UNBOILED at 20 µg

Lane 4:

MCF7 UNBOILED at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 45 kDa

Observed band size: 45 kDa

false