Recombinant Anti-Mesothelin antibody [EPR2685(2)] (ab134109)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR2685(2)] to Mesothelin
- Suitable for: WB, IP, IHC-P
- Reacts with: Human
Related conjugates and formulations
Overview
-
Product name
Anti-Mesothelin antibody [EPR2685(2)]
See all Mesothelin primary antibodies -
Description
Rabbit monoclonal [EPR2685(2)] to Mesothelin -
Host species
Rabbit -
Tested applications
Suitable for: WB, IP, IHC-Pmore details
Unsuitable for: ICC/IF -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human Mesothelin aa 550 to the C-terminus. The exact sequence is proprietary.
Database link: Q13421 -
Positive control
- WB: HeLa whole cell lysate (ab150035); Human fetal lung lysate. OVCAR-3 cell lysate IHC-P: Human mesothelioma and ovarian carcinoma tissue.
-
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. -
Dissociation constant (KD)
KD = 5.50 x 10 -12 M Learn more about KD -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR2685(2) -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
Positive Controls
-
Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab134109 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
---|---|---|
WB |
1/1000 - 1/10000. Detects a band of approximately 40, 69 kDa (predicted molecular weight: 69 kDa).
|
|
IP |
1/10 - 1/100.
|
|
IHC-P |
1/500 - 1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
|
Notes |
---|
WB
1/1000 - 1/10000. Detects a band of approximately 40, 69 kDa (predicted molecular weight: 69 kDa). |
IP
1/10 - 1/100. |
IHC-P
1/500 - 1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Target
-
Function
Membrane-anchored forms may play a role in cellular adhesion.
Megakaryocyte-potentiating factor (MPF) potentiates megakaryocyte colony formation in vitro. -
Tissue specificity
Expressed in lung. Expressed at low levels in heart, placenta and kidney. Expressed in mesothelial cells. Highly expressed in mesotheliomas, ovarian cancers, and some squamous cell carcinomas (at protein level). -
Involvement in disease
Note=Antibodies against MSLN are detected in patients with mesothelioma and ovarian cancer. -
Sequence similarities
Belongs to the mesothelin family. -
Post-translational
modificationsBoth MPF and the cleaved form of mesothelin are N-glycosylated.
Proteolytically cleaved by a furin-like convertase to generate megakaryocyte-potentiating factor (MPF), and the cleaved form of mesothelin. -
Cellular localization
Secreted and Cell membrane. Golgi apparatus. - Information by UniProt
-
Database links
- Entrez Gene: 10232 Human
- Omim: 601051 Human
- SwissProt: Q13421 Human
- Unigene: 408488 Human
-
Alternative names
- CAK 1 antibody
- CAK1 antibody
- CAK1 antigen antibody
see all
Images
-
All lanes : Anti-Mesothelin antibody [EPR2685(2)] (ab134109) at 1/1000 dilution
Lane 1 : OVCAR-3 cell lysate
Lane 2 : HeLa cell lysate
Lane 3 : A549 cell lysate
Lane 4 : PC-3 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 69 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted?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.
-
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.
-
Anti-Mesothelin antibody [EPR2685(2)] (ab134109) at 1/1000 dilution (purified) + HeLa whole cell lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 69 kDa
Observed band size: 69 kDa
Additional bands at: 40 kDa (possible cleavage fragment)Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST -
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
-
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.
-
All lanes : Anti-Mesothelin antibody [EPR2685(2)] (ab134109) at 1/1000 dilution (unpurified)
Lane 1 : HeLa cell lysate
Lane 2 : Human fetal lung lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution
Predicted band size: 69 kDa -
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.
-
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.
-
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.
-
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.
-
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.
Protocols
Datasheets and documents
-
SDS download
-
Datasheet download
References (1)
ab134109 has been referenced in 1 publication.
- Hassan R et al. Clinical Response of Live-Attenuated, Listeria monocytogenes Expressing Mesothelin (CRS-207) with Chemotherapy in Patients with Malignant Pleural Mesothelioma. Clin Cancer Res 25:5787-5798 (2019). PubMed: 31263030