All lanes : Anti-Desmoglein 2/DSG2 antibody [EPR6768] (ab150372) at 1/10000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : DSG2 knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 122 kDa
Observed band size: 90-160 kDa why is the actual band size different from the predicted?

This data was developed using the same antibody clone in a different buffer formulation (ab150372).

Lanes 1- 2: Merged signal (red and green). Green - ab150372 observed at 90-160 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

 ab150372 was shown to react with Desmoglein 2/DSG2 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261826 (knockout cell lysate ab257158) was used. Wild-type HeLa and DSG2 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab150372 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 10000 dilution and a 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.

This IHC data was generated using the same anti-Desmoglein 2/DSG2 antibody clone, EPR6768, in a different buffer formulation (cat# ab150372).

Immunohistochemical analysis of paraffin embedded Human thyroid gland papillary carcinoma tissue labeling Desmoglein 2/DSG2 with ab150372 at a 1/250 dilution.

Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

All lanes : Anti-Desmoglein 2/DSG2 antibody [EPR6768] (ab150372) at 1/10000 dilution

Lane 1 : Trypsinised HAP1 cell lysate
Lane 2 : Scraped HAP1 cell lysate
Lane 3 : Trypsinised DSG2 knockout HAP1 cell lysate
Lane 4 : Scraped DSG2 knockout HAP1 cell lysate
Lane 5 : Trypsinised A431 cell lysate
Lane 6 : Scraped A431 cell lysate

Lysates/proteins at 40 µg per lane.

Performed under reducing conditions.

Predicted band size: 122 kDa
Observed band size: 95 kDa why is the actual band size different from the predicted?

This data was developed using the same antibody clone in a different buffer formulation (ab150372).

Lanes 1 - 6: Merged signal (red and green). Green - ab150372 observed at 95 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab150372 was shown to react with Desmoglein 2 in wild-type HAP1 cells in western blot with loss of signal observed in DSG2 knockout sample. Wild-type and DSG2 knockout HAP1 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab150372 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.