Recombinant Anti-TAGLN/Transgelin antibody [EPR21206] (ab213273)

Immunohistochemical analysis of paraffin-embedded rat esophagus tissue sections labelling TAGLN/Transgelin at 1/100 dilution (5 μg/ml) followed by Goat Anti-Human IgG H&L (HRP) (ab6858) secondary antibody at 1/1000 dilution. Positive staining on the smooth muscle in rat esophagus. The section was incubated with ab213273 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes.

Immunohistochemical analysis of paraffin-embedded rat lung tissue sections labelling TAGLN/Transgelin at 1/100 dilution (5 μg/ml) followed by Goat Anti-Human IgG H&L (HRP) (ab6858) secondary antibody at 1/1000 dilution. Positive staining on the smooth muscle in rat lung. The section was incubated with ab213273 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes.

Immunohistochemical analysis of paraffin-embedded rat skeletal muscle tissue sections labelling TAGLN/Transgelin at 1/100 dilution (5 μg/ml) followed by Goat Anti-Human IgG H&L (HRP) (ab6858) secondary antibody at 1/1000 dilution. Negative control: no staining in rat skeletal muscle. The section was incubated with ab213273 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes.

All lanes : Anti-TAGLN/Transgelin antibody [EPR21206] (ab213273) at 1/500 dilution

Lane 1 : Rat bladder tissue lysate
Lane 2 : Rat colon tissue lysate
Lane 3 : Rat skeletal muscle tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Human IgG Fc (HRP) preadsorbed (ab98624) at 1/10000 dilution

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

Blocking buffer and concentration: 5% NFDM/TBST 

Diluting buffer and concentration: 5% NFDM/TBST

Exposure Time: Lane 1: 3 seconds, Lane 2-3: 20 seconds.

Negative control: Skeletal muscle.

The identity of the bands between 27 kDa and 150 kDa are unknown.

ab181602 was used as a loading control.

All lanes : Anti-TAGLN/Transgelin antibody [EPR21206] (ab213273) at 1 µg/ml

Lane 1 : Human skeletal muscle tissue lysate
Lane 2 : Human colon tissue lysate
Lane 3 : Mouse skeletal muscle tissue lysate
Lane 4 : Mouse Colon tissue lysate
Lane 5 : Mouse Bladder Tissue Lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat anti-Human IgG H&L (IRDye® 800CW) at 1/10000 dilution

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


Exposure time: 3 minutes

This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 40 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab213273 overnight at 4°C. Blots were developed with Goat anti-Human IgG H&L (IRDye^® 800CW) preabsorbed secondary at 1/10000 dilution for 1 hour. 

IHC image of SM22 alpha staining in a section of formalin-fixed paraffin-embedded normal mouse bladder performed on a Leica BOND^TM. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab213273, 5ug/ml, for 15 mins at room temperature.

An HRP-conjugated goat anti-Human IgG secondary was used for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

IHC image of SM22 alpha staining in a section of formalin-fixed paraffin-embedded normal human bladder* performed on a Leica BOND^TM. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab213273, 5ug/ml, for 15 mins at room temperature.

An HRP-conjugated goat anti-Human IgG secondary was used for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre