All lanes : Anti-TAGLN/Transgelin antibody (ab14106) at 1/1000 dilution

All lanes : Lysates prepared from human primary smooth muscle cells

Lysates/proteins at 30 µg per lane.

Secondary
All lanes : HRP-conjugated bovine polyclonal to rabbit IgG at 1/2000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 23 kDa
Observed band size: 23 kDa


Exposure time: 2 minutes

See Abreview

ab14106 stained HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.

The cells were fixed in 100% methanol for 5 minutes, permeabilized in 0.1% PBS-Triton X-100 for 5 minutes and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour at room temperature to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab14106 at 1 µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed used at a 1/1000 dilution for 1 hour at room temperature. Alexa Fluor^® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1 hour at room temperature.

DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1 hour at room temperature.

ab14106 staining SM22 alpha in mouse muscle cells by ICC/IF (Immunocytochemistry/immunofluorescence).

Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 10% serum for 30 minutes at 25°C. Samples were incubated with primary antibody (1/100 in PBS) for 18 hours at 4°C. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody at a dilution of 1/1000.

See Abreview

All lanes : Anti-TAGLN/Transgelin antibody (ab14106) at 1 µg/ml

Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : HeLa nuclear extract
Lane 3 : Human skeletal muscle tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Alexa Fluor Goat polyclonal to Rabbit IgG at 1/10000 dilution

Predicted band size: 23 kDa
Observed band size: 23 kDa

The band was completed abolished by peptide blocking with ab16067 (immunizing peptide) - not shown.

 

 

 

 

All lanes : Anti-TAGLN/Transgelin antibody (ab14106) at 1 µg/ml

Lane 1 : Human colon tissue lysate - total protein (ab30051)
Lane 2 : Mouse colon tissue lysate
Lane 3 : Rat colon tissue lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed at 1/50000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 23 kDa
Observed band size: 23 kDa
Additional bands at: 115 kDa, 20 kDa, 36 kDa. We are unsure as to the identity of these extra bands.


Exposure time: 5 seconds

This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab14106 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

Anti-TAGLN/Transgelin antibody (ab14106) at 1 µg/ml + Recombinant Human TAGLN/Transgelin protein (ab101469) at 0.01 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 23 kDa


Exposure time: 30 seconds