Anti-CD73 antibody [4G6E3] (ab202122)
![Western blot - Anti-CD73 antibody [4G6E3] (ab202122)](https://www.abcam.com/ps/products/202/ab202122/Images/ab202122-483361-anti-cd73-antibody-4g6e3-western-blot-wildtype-a375-nt5e-knockout-a375-a431-hepg2.jpg "Western blot - Anti-CD73 antibody [4G6E3] (ab202122)")
Key features and details
- Mouse monoclonal [4G6E3] to CD73
- Suitable for: Flow Cyt, IHC-P, WB
- Knockout validated
- Reacts with: Human
- Isotype: IgG1
Overview
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Product name
Anti-CD73 antibody [4G6E3]
See all CD73 primary antibodies -
Description
Mouse monoclonal [4G6E3] to CD73 -
Host species
Mouse -
Tested applications
Suitable for: Flow Cyt, IHC-P, WBmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat, Cow
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Immunogen
Recombinant fragment corresponding to Human CD73 aa 1-250. Expressed in E. Coli.
Database link: P21589 -
Positive control
- Flow Cytometry: A431 cells; WB: A375, and A431 cell lysates; IHC-P: Human endometrial cancer and Human esophageal cancer tissues.
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General notes
This product was changed from ascites to supernatant. Lot no’s high than GR287343-13 are from Tissue Culture Supernatant
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.05% Sodium azide
Constituent: 99% PBS -
Concentration information loading... -
Purity
Protein G purified -
Purification notes
Purified from tissue culture supernatant. -
Clonality
Monoclonal -
Clone number
4G6E3 -
Isotype
IgG1 -
Research areas
Associated products
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab202122 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 |
|---|---|---|
| Flow Cyt |
1/200 - 1/400.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
|
| IHC-P | (1) |
1/200 - 1/1000.
|
| WB |
1/500 - 1/2000.
|
| Notes |
|---|
|
Flow Cyt
1/200 - 1/400. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
|
IHC-P
1/200 - 1/1000. |
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WB
1/500 - 1/2000. |
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: 281363 Cow
- Entrez Gene: 4907 Human
- Entrez Gene: 23959 Mouse
- Entrez Gene: 58813 Rat
- Omim: 129190 Human
- SwissProt: Q05927 Cow
- SwissProt: P21589 Human
- SwissProt: Q61503 Mouse
see all -
Alternative names
- 5' NT antibody
- 5' nucleotidase (CD73) antibody
- 5' nucleotidase precursor antibody
see all
Images
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Western blot - Anti-CD73 antibody [4G6E3] (ab202122)All lanes : Anti-CD73 antibody [4G6E3] (ab202122) at 1/500 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.
Observed band size: 65 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-CD73 antibody [4G6E3] staining at 1/500 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab202122 was shown to bind specifically to CD73. A band was observed at 65-70 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). A cross-reactive band is observed at 65 kDa in all samples. 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 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-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.
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![Flow Cytometry - Anti-CD73 antibody [4G6E3] (ab202122)](https://www.abcam.com/ps/products/202/ab202122/Images/ab202122-243400-anti-cd73-antibody-4g6e3-flow-cytometry.jpg)
Flow Cytometry - Anti-CD73 antibody [4G6E3] (ab202122)Flow cytometric analysis of A431 cells using ab202122 at 1/200 (green) and negative control (red).
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD73 antibody [4G6E3] (ab202122)](https://www.abcam.com/ps/products/202/ab202122/Images/ab202122-243397-anti-cd73-antibody-4g6e3-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD73 antibody [4G6E3] (ab202122)Immunohistochemical analysis of paraffin-embedded Human esophageal cancer tissue labeling CD73 with ab202122 at 1/200 with DAB staining.
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![Western blot - Anti-CD73 antibody [4G6E3] (ab202122)](https://www.abcam.com/ps/products/202/ab202122/Images/ab202122-353576-anticd73antibody4g6e3westernblotknockouta431nt5ea375humanlysates.jpg)
Western blot - Anti-CD73 antibody [4G6E3] (ab202122)All lanes : Anti-CD73 antibody [4G6E3] (ab202122) at 1/1000 dilution
Lane 1 : Wild-type A-431 whole cell lysate
Lane 2 : NT5E knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 3 : A375 (Human malignant melanoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.Lanes 1 - 3: Merged signal (red and green). Green - ab202122 observed at 63 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab202122 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. Ab202122 and ab181602 (Rabbit 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-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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![Western blot - Anti-CD73 antibody [4G6E3] (ab202122)](https://www.abcam.com/ps/products/202/ab202122/Images/ab202122-243394-anti-cd73-antibody-4g6e3-western-blot.jpg)
Western blot - Anti-CD73 antibody [4G6E3] (ab202122)Anti-CD73 antibody [4G6E3] (ab202122) at 1/500 dilution + A431 cell lysate -
![Western blot - Anti-CD73 antibody [4G6E3] (ab202122)](https://www.abcam.com/ps/products/202/ab202122/Images/ab202122-243395-anti-cd73-antibody-4g6e3-western-blot.jpg)
Western blot - Anti-CD73 antibody [4G6E3] (ab202122)Anti-CD73 antibody [4G6E3] (ab202122) at 1/500 dilution + Human CD73 (AA: 30-250) recombinant proteinExpected MW is 26.6 kDa
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD73 antibody [4G6E3] (ab202122)](https://www.abcam.com/ps/products/202/ab202122/Images/ab202122-243396-anti-cd73-antibody-4g6e3-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD73 antibody [4G6E3] (ab202122)Immunohistochemical analysis of paraffin-embedded Human endometrial cancer tissue labeling CD73 with ab202122 at 1/200 with DAB staining.
Protocols
Datasheets and documents
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Datasheet download
References (3)
ab202122 has been referenced in 3 publications.
- Li B et al. The MSC-Derived Exosomal lncRNA H19 Promotes Wound Healing in Diabetic Foot Ulcers by Upregulating PTEN via MicroRNA-152-3p. Mol Ther Nucleic Acids 19:814-826 (2020). PubMed: 31958697
- Pan C et al. Pretreatment with human urine-derived stem cells protects neurological function in rats following cardiopulmonary resuscitation after cardiac arrest. Exp Ther Med 20:112 (2020). PubMed: 32989390
- Yu L et al. Exosomes derived from microRNA-199a-overexpressing mesenchymal stem cells inhibit glioma progression by down-regulating AGAP2. Aging (Albany NY) 11:5300-5318 (2019). PubMed: 31386624
