Recombinant Anti-Mesothelin antibody [EPR19025-42] - BSA and Azide free (ab227810)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19025-42] to Mesothelin - BSA and Azide free
- Suitable for: WB, IHC-P, Flow Cyt, IP
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Mesothelin antibody [EPR19025-42] - BSA and Azide free
See all Mesothelin primary antibodies -
Description
Rabbit monoclonal [EPR19025-42] to Mesothelin - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, Flow Cyt, IPmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- IHC-P: Human mesothelioma tissue. WB: OVCAR-3 cell lysate. Flow cyt: HeLa cells
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General notes
ab227810 is the carrier-free version of ab196235.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR19025-42 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab227810 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 |
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WB |
Use at an assay dependent concentration. Detects a band of approximately 40, 69 kDa (predicted molecular weight: 69 kDa).
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Flow Cyt |
Use at an assay dependent concentration.
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IP |
Use at an assay dependent concentration.
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Notes |
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WB
Use at an assay dependent concentration. Detects a band of approximately 40, 69 kDa (predicted molecular weight: 69 kDa). |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Flow Cyt
Use at an assay dependent concentration. |
IP
Use at an assay dependent concentration. |
Target
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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
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Database links
- Entrez Gene: 10232 Human
- Omim: 601051 Human
- SwissProt: Q13421 Human
- Unigene: 408488 Human
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Alternative names
- CAK 1 antibody
- CAK1 antibody
- CAK1 antigen antibody
see all
Images
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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab196235).
Flow cytometry overlay histogram showing left HeLa positive cells and right negative PC3 stained with ab196235 (red line). The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interactionfollowed by the antibody (ab196235) (1x 106 in 100μl at 5.0 μg/ml (1/410)) for 30min on ice.
The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min on ice
Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
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All lanes : Anti-Mesothelin antibody [EPR19025-42] (ab196235) 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?This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab196235).
False colour image of Western blot: Anti-Mesothelin antibody [EPR19025-42] 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, ab196235 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.
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Flow cytometric analysis of HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling Mesothelin with ab196235 at 1/50 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
Total viable cells were gated for the image.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab196235).
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Immunohistochemical analysis of paraffin-embedded human ovarian adenocarcinoma tissue labeling Mesothelin with ab196235 at 1/1500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on tumor cells of human ovarian adenocarcinoma (PMID: 17945478). Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab196235).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Mesothelin was immunoprecipitated from 0.35 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) lysate with ab196235 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab196235 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: HeLa whole cell lysate 10 µg (Input).
Lane 2: ab196235 IP in HeLa whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab196235 in HeLa whole cell lysate.
Exposure time: 3 minutes.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab196235).
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Immunohistochemical analysis of paraffin-embedded human mesothelioma tissue labeling Mesothelin with ab196235 at 1/1500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and cytoplasmic staining on human mesothelioma (PMID: 17945478). Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab196235).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab227810 has not yet been referenced specifically in any publications.