Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling Liver Arginase with ab233548 at 1/2000 dilution (0.333 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear and cytoplasmic staining in rat liver (PMID/ 22298472, 23637937, 21975054) is observed. The section was incubated with ab233548 for 15 mins at RT. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

All lanes : Anti-Liver Arginase antibody [EPR22033-369] (ab233548) at 1/1000 dilution

Lane 1 : Mouse liver tissue lysate
Lane 2 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 35 kDa
Observed band size: 37 kDa why is the actual band size different from the predicted?


Exposure time: 1 second

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

The expression profile observed in RAW 264.7 is consistent with the literature (PMID: 28032600, PMID: 27166374, PMID: 9814991, PMID: 11080091).

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Hepa1-6 (mouse epithelial hepatoma) cells labeling Liver Arginase with ab233548 at 1/60 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730, Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

Liver Arginase was immunoprecipitated from 0.35 mg Mouse liver tissue lysate with ab233548 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab233548 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1: Mouse liver tissue lysate 10ug

Lane 2: ab233548 IP in Mouse liver tissue lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab233548 in Mouse liver tissue lysate

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

Exposure time: 3 seconds

 

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Liver Arginase with ab233548 at 1/2000 dilution (0.333 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear and cytoplasmic staining in mouse liver (PMID/ 22298472, 23637937, 21975054). The section was incubated with ab233548 for 15 mins at RT. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control/ Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

All lanes : Anti-Liver Arginase antibody [EPR22033-369] (ab233548) at 1/5000 dilution

Lane 1 : Mouse liver lysate
Lane 2 : Mouse kidney lysate
Lane 3 : Rat liver lysate
Lane 4 : Rat kidney lysate

Lysates/proteins at 20 µg per lane.

Secondary
Lanes 1-2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Lanes 3-4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 35 kDa
Observed band size: 37 kDa why is the actual band size different from the predicted?

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 27761413).   Negative control: kidney (PMID: 21975054)

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

Exposure Time: 1 second