Anti-Liver Arginase antibody [EPR22033-369]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Liver Arginase antibody [EPR22033-369] (AB233548)

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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Liver Arginase antibody [EPR22033-369] (AB233548)

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.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Liver Arginase antibody [EPR22033-369] (AB233548)

Intracellular 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.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Liver Arginase antibody [EPR22033-369] (AB233548)

Flow cytometry staining of C57BL/6 Mouse Bone Marrow Cell-derived Macrophages (M1 polarisation) (top) or C57BL/6 Mouse Bone Marrow Cell-derived Macrophages (M2 polarisation) (bottom) with ab233548 (right) or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (left). Cells were fixed and permeabilised with BD Cytofix/Cytoperm™ for 20 min. Cells were incubated for 30min on ice in 1x PBS containing 10μg/ml anti CD16/CD32 and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab233548 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 10⁶in 100 μl at 5.0 μg/ml (1/472)) for 30min at 22°C. The cells were simultaneously stained with F4/80.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on live cells.

• IP

Unknown

Immunoprecipitation - Anti-Liver Arginase antibody [EPR22033-369] (AB233548)

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

All lanes:

Immunoprecipitation - Anti-Liver Arginase antibody [EPR22033-369] (ab233548)

Predicted band size: 35 kDa

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• WB

Lab

Western blot - Anti-Liver Arginase antibody [EPR22033-369] (AB233548)

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).

All lanes:

Western blot - Anti-Liver Arginase antibody [EPR22033-369] (ab233548) at 1/1000 dilution

Lane 1:

Mouse liver tissue lysate at 20 µg

Lane 2:

RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 35 kDa

Observed band size: 37 kDa

false

Exposure time: 1s

• WB

Lab

Western blot - Anti-Liver Arginase antibody [EPR22033-369] (AB233548)

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

All lanes:

Western blot - Anti-Liver Arginase antibody [EPR22033-369] (ab233548) at 1/5000 dilution

Lane 1:

Mouse liver lysate at 20 µg

Lane 2:

Mouse kidney lysate at 20 µg

Lane 3:

Rat liver lysate at 20 µg

Lane 4:

Rat kidney lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 35 kDa

Observed band size: 37 kDa

false