Recombinant Anti-CD73 antibody [EPR6114] (ab133582)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR6114] to CD73
- Suitable for: IHC-P, WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-CD73 antibody [EPR6114]
See all CD73 primary antibodies -
Description
Rabbit monoclonal [EPR6114] to CD73 -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, WBmore details
Unsuitable for: Flow Cyt,ICC/IF or IP -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human fetal liver tissue lysate, Human liver, Human spleen; A375 and HepG2 whole cell lysate (ab7900) IHC-P: Human lung carcinoma, Human tonsil tissue
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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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
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
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, PBS, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR6114 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab133582 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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IHC-P |
1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
For unpurified use ad 1/100 - 1/250. See IHC antigen retrieval protocols. |
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WB | (1) |
1/1000 - 1/10000. Predicted molecular weight: 63 kDa.
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Notes |
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IHC-P
1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. For unpurified use ad 1/100 - 1/250. See IHC antigen retrieval protocols. |
WB
1/1000 - 1/10000. Predicted molecular weight: 63 kDa. |
Target
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Function
Hydrolyzes extracellular nucleotides into membrane permeable nucleosides. -
Involvement in disease
Defects in NT5E are the cause of calcification of joints and arteries (CAJA) [MIM:211800]. A condition characterized by adult-onset calcification of the lower extremity arteries, including the iliac, femoral and tibial arteries, and hand and foot capsule joints. Age of onset has been reported as early as the second decade of life, usually involving intense joint pain or calcification in the hands. -
Sequence similarities
Belongs to the 5'-nucleotidase family. -
Cellular localization
Cell membrane. - Information by UniProt
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Database links
- Entrez Gene: 4907 Human
- Omim: 129190 Human
- SwissProt: P21589 Human
- Unigene: 153952 Human
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Alternative names
- 5' NT antibody
- 5' nucleotidase (CD73) antibody
- 5' nucleotidase precursor antibody
see all
Images
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All lanes : Anti-CD73 antibody [EPR6114] (ab133582) at 1/1000 dilution
Lane 1 : Wild-type A-375 cell lysate
Lane 2 : NT5E knockout A-375 cell lysate
Lane 3 : A431 cell lysate
Lane 4 : HepG2 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 63 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-CD73 antibody [EPR6114] 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, ab133582 was shown to bind specifically to CD73. A band was observed at 70 kDa in wild-type A-375 cell lysates with no signal observed at this size in NT5E knockout cell line ab281591 (knockout cell lysate ab282943). To generate this image, wild-type and NT5E knockout A-375 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution 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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All lanes : Anti-CD73 antibody [EPR6114] (ab133582) at 1/1000 dilution
Lane 1 : Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 2 : NT5E knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 3 : A-375 (Human malignant melanoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 63 kDaLanes 1 - 3: Merged signal (red and green). Green - ab133582 observed at 63 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab133582 was shown to recognize NT5E in wild-type A-431 cells as signal was lost at the expected MW in NT5E knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and NT5E knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab133582 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD73 antibody [EPR6114] (ab133582)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue sections labeling CD73 with Purified ab133582 at 1:2000 dilution (1.12 µg/ml).
Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0).
ImmunoHistoProbe one step HRP Polymer (ready to use) was used.
Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD73 antibody [EPR6114] (ab133582)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue sections labeling CD73 with Purified ab133582 at 1:2000 dilution (1.12 µg/ml).
Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0).
ImmunoHistoProbe one step HRP Polymer (ready to use)was used.
Negative control:PBS instead of the primary antibody.
Hematoxylin was used as a counterstain.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD73 antibody [EPR6114] (ab133582)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue sections labeling CD73 with Purified ab133582 at 1:2000 dilution (1.12 µg/ml).
Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0).
ImmunoHistoProbe one step HRP Polymer (ready to use) was used.
Negative control: PBS instead of the primary antibody.
Hematoxylin was used as a counterstain.
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Anti-CD73 antibody [EPR6114] (ab133582) at 1/1000 dilution + A375 ( Human malignant melanoma epithelial cell) whole cell lysates at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 63 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD73 antibody [EPR6114] (ab133582)
Immunohistochemical analysis of paraffin-embedded human tonsil tissue labelling CD73 with ab133582 at 1/100 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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All lanes : Anti-CD73 antibody [EPR6114] (ab133582) at 1/1000 dilution
Lane 1 : Human fetal liver tissue lysate
Lane 2 : A375 cell lysate
Lane 3 : HepG2 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled Goat anti-Rabbit IgG at 1/2000 dilution
Predicted band size: 63 kDa
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (15)
ab133582 has been referenced in 15 publications.
- Hara T et al. CD73+ Epithelial Progenitor Cells That Contribute to Homeostasis and Renewal Are Depleted in Eosinophilic Esophagitis. Cell Mol Gastroenterol Hepatol 13:1449-1467 (2022). PubMed: 35108658
- Liu C et al. Dental pulp stem cell-derived exosomes suppress M1 macrophage polarization through the ROS-MAPK-NFκB P65 signaling pathway after spinal cord injury. J Nanobiotechnology 20:65 (2022). PubMed: 35109874
- Zhou Y et al. High expression of CD73 contributes to poor prognosis of clear-cell renal cell carcinoma by promoting cell proliferation and migration. Transl Cancer Res 11:3634-3644 (2022). PubMed: 36388013
- Liu H et al. Human amnion-derived mesenchymal stem cells improved the reproductive function of age-related diminished ovarian reserve in mice through Ampk/FoxO3a signaling pathway. Stem Cell Res Ther 12:317 (2021). PubMed: 34078462
- Vos H et al. Comparison of the tumor immune microenvironment of primary hormone receptor-negative HER2-positive and triple negative breast cancer. NPJ Breast Cancer 7:128 (2021). PubMed: 34556657