Anti-Liver Arginase antibody [EPR6672(B)]

• IHC-P

Lab

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 (ab209101). The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• IHC-P

Unknown

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 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.

• IP

Lab

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 (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) (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

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Exposure time: 1s

• WB

Lab

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 1/20000 dilution

All lanes:

Human liver lysates at 15 µg

Secondary

All lanes:

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

Predicted band size: 35 kDa

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Exposure time: 5s

• WB

Lab

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] (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 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (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

Lane 2:

Western blot - Human ARG1 knockout Hep G2 cell line (<a href='/ko/products/cell-lines/human-arg1-knockout-hep-g2-cell-line-ab281603'>ab281603</a>)

Lanes 3 and 5:

Empty

Lane 4:

Human Liver cell lysate at 5 µg

Lane 6:

A549 cell lysate at 20 µg

Predicted band size: 35 kDa

Observed band size: 36 kDa

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• WB

Unknown

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) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 35 kDa

Observed band size: 61 kDa

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• WB

CiteAb

Western blot - Anti-Liver Arginase antibody [EPR6672(B)] (AB133543)

Western Blotting using Anti-Liver Arginase antibody [EPR6672(B)], ab133543. Publication image from Lin, L. et al., 2017, Nat Commun, 28808255. Legend direct from paper.

Glutamine nitrogens do not integrate into the urea cycle in vitro. a Immunoblots for ARG2 and ARG1 in AsPC1-shScr and AspC1-shARG2 cells. b Proliferation curves of cells in a (n = 6). Data are representative of 4 independent experiments. c Fractional 13C-labeling of urea (M1) and ornithine (M5) in AsPC-1 cells with control (shScr) or ARG2 knockdown (shARG2, hairpins A06 and A10) grown in culture in 0.1% serum and 100 ng ml−1 IGF-1, supplemented with 1 mM 13C6-arginine for 48 h (n = 4). d Diagram of the urea cycle, with metabolites in red indicating significant 15N-labeling. e Fractional labeling of the indicated metabolites in cells described in a, grown in media supplemented with 2 mM 15N(amine)-glutamine for 1 h or 24 h (n = 4). Data are representative of two independent experiments. In b, data represent the mean ± s.e.m. In c, e, data represent the mean ± s.d. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001, by one-way ANOVA followed by Tukey test

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