Anti-N-Cadherin/CD325 antibody [32/N-Cadherin]

• IP

Supplier Data

Immunoprecipitation - Anti-N-Cadherin/CD325 antibody [32/N-Cadherin] (AB280375)

N Cadherin was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab280375 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab280375 at 1/1000 dilution. HRP-conjuagted mouse IgG for IP (ab131368) was used at 1/1000 dilution.

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug

Lane 2 : ab280375 IP in HeLa whole cell lysate

Lane 3 : Mouse monoclonal IgG1 (ab18443) instead of ab280375 in HeLa whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 23 seconds

The molecular weight observed is consistent with what has been described in the literature (PMID : 22553038)

All lanes:

Immunoprecipitation - Anti-N-Cadherin/CD325 antibody [32/N-Cadherin] (ab280375)

Predicted band size: 99 kDa

false

• WB

Supplier Data

Western blot - Anti-N-Cadherin/CD325 antibody [32/N-Cadherin] (AB280375)

Blocking and Diluting buffer and concentration : 5% NFDM/TBST

Exposure time 26 seconds

Negative control : MCF7 (PMID : 9177902)

All lanes:

Western blot - Anti-N-Cadherin/CD325 antibody [32/N-Cadherin] (ab280375) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 10 µg

Lane 2:

MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate at 10 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Predicted band size: 99 kDa

Observed band size: 140 kDa

false

• WB

Supplier Data

Western blot - Anti-N-Cadherin/CD325 antibody [32/N-Cadherin] (AB280375)

Blocking and Diluting buffer and concentration : 5% NFDM/TBST

Exposure time 15 seconds

Lanes 1-2 used the fresh lysates to minimize protein degradation.

All lanes:

Western blot - Anti-N-Cadherin/CD325 antibody [32/N-Cadherin] (ab280375) at 1/5000 dilution

Lane 1:

HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate at 10 µg

Lane 2:

293T (human embryonic kidney epithelial cell), whole cell lysate at 10 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Predicted band size: 99 kDa

Observed band size: 140 kDa

false

• WB

Supplier Data

Western blot - Anti-N-Cadherin/CD325 antibody [32/N-Cadherin] (AB280375)

Blocking and Diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.

Lanes 1 - 2 : Merged signal (red and green). Green - ab280375 observed at 130 kDa. Red - loading control ab181602(Rabbit monoclonal [EPR16891] to GAPDH) observed at 36 kDa.

Lanes 1-2 : ab280375 Anti-CDH2 antibody was shown to react with CDH2 in 293T cells in Western blot. Loss of signal was observed when CDH2 knockout sample was used. Wild-type and CDH2 knockout samples were subjected to SDS-PAGE.

ab280375 and Anti-GAPDH antibody [EPR16891] (ab181602) were incubated at 4℃ overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680CW) (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800RD) (ab216772) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-N-Cadherin/CD325 antibody [32/N-Cadherin] (ab280375) at 1/1000 dilution

Lane 1:

Wild-type 293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg

Lane 2:

CDH2 knockout 293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Mouse IgG H&L (IRDye® 800CW) (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-800cw-preadsorbed-ab216772'>ab216772</a>) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/10000 dilution

Predicted band size: 99 kDa

Observed band size: 140 kDa

false

• WB

Supplier Data

Western blot - Anti-N-Cadherin/CD325 antibody [32/N-Cadherin] (AB280375)

Blocking and Diluting buffer and concentration : 5% NFDM/TBST

Exposure time Lanes 1-2 : 15 seconds

Lanes 3 : 10 seconds

Lanes 4-5 : 26 seconds

All lanes:

Western blot - Anti-N-Cadherin/CD325 antibody [32/N-Cadherin] (ab280375) at 1/1000 dilution

Lane 1:

Mouse brain lysate at 10 µg

Lane 2:

Mouse kidney lysate at 10 µg

Lane 3:

Rat heart lysate at 10 µg

Lane 4:

C6 (rat glial tumor glial cell), whole cell lysate at 10 µg

Lane 5:

PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate at 10 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Predicted band size: 99 kDa

Observed band size: 140 kDa

false