Anti-Mesothelin antibody [EPR2685(2)]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mesothelin antibody [EPR2685(2)] (AB134109)

Immunohistochemical analysis of paraffin embedded Human ovarian carcinoma tissue labelling Mesothelin with unpurified ab134109 at 1/1000.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mesothelin antibody [EPR2685(2)] (AB134109)

Immunohistochemical staining of paraffin embedded human skeletal muscle with purified ab134109 at a working dilution of 1/800. The secondary antibody used is ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mesothelin antibody [EPR2685(2)] (AB134109)

Immunohistochemical analysis of paraffin embedded Human Greator omentum tissue using unpurified ab134109 showing +ve staining.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mesothelin antibody [EPR2685(2)] (AB134109)

Immunohistochemical analysis of paraffin embedded normal Human pancreas tissue using unpurified ab134109 showing -ve staining.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mesothelin antibody [EPR2685(2)] (AB134109)

Immunohistochemical analysis of paraffin embedded Human mesothelioma tissue labelling Mesothelin with unpurified ab134109 at 1/1000.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mesothelin antibody [EPR2685(2)] (AB134109)

Immunohistochemical analysis of paraffin embedded Human Pancreatic carcinoma tissue using unpurified ab134109 showing +ve staining.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mesothelin antibody [EPR2685(2)] (AB134109)

Immunohistochemical staining of paraffin embedded human mesothelioma with purified ab134109 at a working dilution of 1/800. The secondary antibody used is ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

• IP

Unknown

Immunoprecipitation - Anti-Mesothelin antibody [EPR2685(2)] (AB134109)

ab134109 (purified) at 1/40 immunoprecipitating mesothelin in 10 μg HeLa cell lysate (Lanes 1 and 2, observed at 40 and 69 kDa). Lane 3 - Rabbit monoclonal IgG (ab172730). For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution. Blocking buffer and concentration : 5% NFDM/TBST Dilution buffer and concentration : 5% NFDM/TBST

All lanes:

Immunoprecipitation - Anti-Mesothelin antibody [EPR2685(2)] (ab134109)

Predicted band size: 69 kDa

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• WB

Lab

Western blot - Anti-Mesothelin antibody [EPR2685(2)] (AB134109)

Blocking buffer : 5% NFDM/TBST
Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Mesothelin antibody [EPR2685(2)] (ab134109) at 1/1000 dilution

All lanes:

HeLa whole cell lysate at 10 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: 42 kDa,46 kDa,69 kDa,79 kDa

Observed band size: 40 kDa,42 kDa,69 kDa,79 kDa

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• WB

Lab

Western blot - Anti-Mesothelin antibody [EPR2685(2)] (AB134109)

False colour image of Western blot : Anti-Mesothelin antibody [EPR2685(2)] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab134109 was shown to bind specifically to Mesothelin. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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-Mesothelin antibody [EPR2685(2)] (ab134109) at 1/1000 dilution

Lane 1:

OVCAR-3 cell lysate at 20 µg

Lane 2:

HeLa cell lysate at 20 µg

Lane 3:

A549 cell lysate at 20 µg

Lane 4:

PC-3 cell lysate at 20 µg

Predicted band size: 69 kDa

Observed band size: 45 kDa

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• WB

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Western blot - Anti-Mesothelin antibody [EPR2685(2)] (AB134109)

All lanes:

Western blot - Anti-Mesothelin antibody [EPR2685(2)] (ab134109) at 1/1000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

Human fetal lung lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution

Predicted band size: 17 kDa,19 kDa,36 kDa,69 kDa

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• OI-RD Scanning

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OI-RD Scanning - Anti-Mesothelin antibody [EPR2685(2)] (AB134109)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.