Anti-Liver Arginase antibody [EPR6672(B)] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Liver Arginase antibody [EPR6672(B)] - BSA and Azide free (AB211961)

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. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133543).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Liver Arginase antibody [EPR6672(B)] - BSA and Azide free (AB211961)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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)] - BSA and Azide free (AB211961)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133543).

All lanes:

Immunoprecipitation - Anti-Liver Arginase antibody [EPR6672(B)] (<a href='/en-us/products/primary-antibodies/liver-arginase-antibody-epr6672b-ab133543'>ab133543</a>)

Predicted band size: 35 kDa

false

Exposure time: 1s

• WB

Lab

Western blot - Anti-Liver Arginase antibody [EPR6672(B)] - BSA and Azide free (AB211961)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133543).

All lanes:

Western blot - Anti-Liver Arginase antibody [EPR6672(B)] (<a href='/en-us/products/primary-antibodies/liver-arginase-antibody-epr6672b-ab133543'>ab133543</a>) 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='/en-us/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

false

• WB

Lab

Western blot - Anti-Liver Arginase antibody [EPR6672(B)] - BSA and Azide free (AB211961)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133543).

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Liver Arginase antibody [EPR6672(B)] (<a href='/en-us/products/primary-antibodies/liver-arginase-antibody-epr6672b-ab133543'>ab133543</a>) 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='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 35 kDa

false

Exposure time: 5s

• WB

Unknown

Western blot - Anti-Liver Arginase antibody [EPR6672(B)] - BSA and Azide free (AB211961)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133543).

All lanes:

Western blot - Anti-Liver Arginase antibody [EPR6672(B)] (<a href='/en-us/products/primary-antibodies/liver-arginase-antibody-epr6672b-ab133543'>ab133543</a>) 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='/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: 61 kDa

false