Anti-Liver Arginase antibody [EPR6672(B)]

6 product images

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  Western blot - Anti-Liver Arginase antibody [EPR6672(B)] (ab133543)

  False colour image of Western blot: Anti-Liver Arginase antibody [EPR6672(B)] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab133543 was shown to bind specifically to Liver Arginase. A band was observed at 36 kDa in wild-type HepG2 cell lysates with no signal observed at this size in arg1 knockout cell line Human ARG1 knockout Hep G2 cell line ab281603 (knockout cell lysate ab282955). To generate this image, wild-type and arg1 knockout HepG2 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.

  All lanes: Western blot - Anti-Liver Arginase antibody [EPR6672(B)] (ab133543) at 1/1000 dilution

  Lane 1: Wild-type HepG2 cell lysate at 20 µg

  Lane 2: arg1 knockout HepG2 cell lysate at 20 µg

  Lanes 3 and 5: Empty

  Lane 4: Human Liver cell lysate at 5 µg

  Lane 6: A549 cell lysate at 20 µg

  Performed under reducing conditions.

  Predicted band size: 35 kDa

  Observed band size: 36 kDa

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  Western blot - Anti-Liver Arginase antibody [EPR6672(B)] (ab133543)

  All lanes: Western blot - Anti-Liver Arginase antibody [EPR6672(B)] (ab133543) at 1/1000 dilution

  Lane 1: GST tagged Recombinant Human ARG1 protein (full-length, aa 1 to 322) (61 KDa)

  Lane 2: GST tagged Recombinant Human ARG2 protein (full-length, aa 1 to 354) (65 KDa）

  Secondary

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

  Predicted band size: 35 kDa

  Observed band size: 61 kDa

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  Immunoprecipitation - Anti-Liver Arginase antibody [EPR6672(B)] (ab133543)

  Lane 1 (input): Human fetal liver whole cell lysate, 10μg
  Lane 2: Human fetal liver whole cell lysate
  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab133543 in Human fetal liver whole cell lysate
  

  ab133543 immunoprecipitating liver arginase in Human fetal liver whole cell lysates. Primary antibody was used at a 1:1000 dilution (0.27 μg/ml). For western blotting, VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used for detection at 1:5000 dilution. Capture antibody was used at 1:20 dilution (1.3μg in 0.35mg lysates).

  Blocking and diluting buffer used: 5% NFDM/TBST.

  All lanes: Immunoprecipitation - Anti-Liver Arginase antibody [EPR6672(B)] (ab133543)

  Predicted band size: 35 kDa

  Exposure time: 1s

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  Western blot - Anti-Liver Arginase antibody [EPR6672(B)] (ab133543)

  Blocking and diluting buffer: 5% NFDM/TBST.

  All lanes: Western blot - Anti-Liver Arginase antibody [EPR6672(B)] (ab133543) at 1/2000 dilution

  All lanes: Human liver lysates at 15 µg

  Secondary

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

  Predicted band size: 35 kDa

  Exposure time: 5s

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Liver Arginase antibody [EPR6672(B)] (ab133543)

  ab133543 staining liver arginase in paraffin embedded human hepatocellular cancer tissue sections by Immunohistochemistry.
  

  Heat mediated antigen retrieval was performed using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).

  Samples were incubated with primary antibody at 1:2000 dilution (0.13 μg/ml).

  A ready to use Goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody.

  Hematoxylin was used as a counterstain.

  Cytoplasmic and nuclear staining on human hepatocellular cancer.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Liver Arginase antibody [EPR6672(B)] (ab133543)

  Tissue Microarrays stained for " Anti-Liver Arginase antibody [EPR6672(B)]” using " ab133543" in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. The sections were incubated with ab133543 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.