Anti-CD73 antibody [RM2050] - BSA and Azide free (ab317463)

Knockout Tested Rabbit Recombinant Multiclonal CD73 antibody. Carrier free. Suitable for WB, Flow Cyt, IP, ICC/IF, IHC-P and reacts with Human, Mouse samples.

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Images

Flow Cytometry - Anti-CD73 antibody [RM2050] - BSA and Azide free (AB317463), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: 100% PBS
Form: Liquid
Clonality: Multiclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	WB	Flow Cyt	IP	ICC/IF	IHC-P
Human	Tested	Tested	Tested	Tested	Tested
Mouse	Tested	Tested	Tested	Tested	Not recommended
Rat	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Human, Mouse	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human, Mouse	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human, Mouse	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human, Mouse	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info -	Notes -

Target data

See full target information NT5E

Function

Catalyzes the hydrolysis of nucleotide monophosphates, releasing inorganic phosphate and the corresponding nucleoside, with AMP being the preferred substrate (PubMed:21933152, PubMed:22997138, PubMed:23142347, PubMed:24887587, PubMed:34403084). Shows a preference for ribonucleotide monophosphates over their equivalent deoxyribose forms (PubMed:34403084). Other substrates include IMP, UMP, GMP, CMP, dAMP, dCMP, dTMP, NAD and NMN (PubMed:21933152, PubMed:22997138, PubMed:23142347, PubMed:24887587, PubMed:34403084).

Alternative names

CD73, NT5, NTE, NT5E, 5'-nucleotidase, 5'-NT, 5'-deoxynucleotidase, Ecto-5'-nucleotidase, IMP-specific 5'-nucleotidase, Thymidylate 5'-phosphatase

Recommended products

Knockout Tested Rabbit Recombinant Multiclonal CD73 antibody. Carrier free. Suitable for WB, Flow Cyt, IP, ICC/IF, IHC-P and reacts with Human, Mouse samples.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: 100% PBS
Form: Liquid
Clonality: Multiclonal
Immunogens: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
Clone number: RM2050
Purification technique: Affinity purification Protein A
Specificity: Unsuitable for mouse IHC.
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: +4°C

Notes

ab317463 is the carrier-free version of Anti-CD73 antibody [RM2050] ab317462.

This product is a recombinant multiclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

CD73 also known as ecto-5'-nucleotidase is an enzyme that functions at the cellular level. The CD73 protein hydrolyzes extracellular nucleotides into nucleosides by removing phosphate groups. It is a 70 kDa glycosylphosphatidylinositol-anchored molecule found on the surface of various cell types including endothelial cells lymphocytes and epithelial cells. Its presence is noted in a wide range of tissues across the human body highlighting its versatile role in cellular communication and metabolism.

Biological function summary

CD73 participates in the regulation of purinergic signaling through adenosine production. It acts in cooperation with other cell surface enzymes such as CD39 forming a functional complex that handles ATP breakdown to adenosine. This enzymatic activity modulates immune responses tissue protection and inflammation control. In addition to immune-related functions CD73 supports the maintenance of vascular integrity and proper cellular adhesion which is important in different physiological and pathological contexts.

Pathways

CD73 plays an important part in the adenosinergic pathway governing the conversion of AMP into adenosine. This activity impacts the signaling and function of the adenosine receptors on immune and non-immune cells. Another important pathway is the angiogenesis process where CD73 contributes by affecting endothelial cell migration and blood vessel development. Relationships with key proteins such as adenosine A2 receptors help facilitate these critical pathway activities.

Associated diseases and disorders

CD73's involvement with immunosuppression and tumor progression becomes significant. Elevated levels of CD73 are frequently observed in some cancers where it aids in creating a favorable tumor microenvironment through increased adenosine production. This situation assists tumor cells in evading immune detection. Additionally CD73 is linked to autoimmune diseases like multiple sclerosis where it contributes to modulated lymphocyte activity. Connections with proteins such as TGF-beta further illustrate its role in disease-related pathways creating potential targets for therapeutic interventions through the use of anti-CD73 antibodies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

13 product images

Flow Cytometry - Anti-CD73 antibody [RM2050] - BSA and Azide free (ab317463)
This data was developed using Anti-CD73 antibody [RM2050] ab317462, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of Mouse spleen cells labelling CD73 with Anti-CD73 antibody [RM2050] ab317462 at 1/50 dilution (1 ug)/Right compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.
Mouse spleen cell are co-stained with CD3 conjugated Alexa Fluor®647.

Gated on viable cell.
Flow Cytometry - Anti-CD73 antibody [RM2050] - BSA and Azide free (ab317463)
This data was developed using Anti-CD73 antibody [RM2050] ab317462, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of HeLa(human cervical adenocarcinoma epithelial cell, Left) / U-87 MG(human glioblastoma-astrocytoma epithelial cell, Right) cells labelling CD73 with Anti-CD73 antibody [RM2050] ab317462 at 1/5000 dilution (0.01 ug)/Red compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.
Low expression: HeLa (PMID: 8566797)

Gated on viable cell.
Flow Cytometry - Anti-CD73 antibody [RM2050] - BSA and Azide free (ab317463)
This data was developed using Anti-CD73 antibody [RM2050] ab317462, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of CD73 knockout A431(huam CD73 knock out human epidermoid carcinoma epithelial cell, Left) / Parental A431(Right) cells labelling CD73 with Anti-CD73 antibody [RM2050] ab317462 at 1/500 dilution (0.1 ug)/Red compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.
Gated on viable cell.
Flow Cytometry - Anti-CD73 antibody [RM2050] - BSA and Azide free (ab317463)
This data was developed using Anti-CD73 antibody [RM2050] ab317462, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of RAW 264.7(mouse Abelson murine leukemia virus-induced tumor macrophage, Left) / 4T1 (mouse mammary gland carcinoma epithelial cell, Right) cells labelling CD73 with Anti-CD73 antibody [RM2050] ab317462 at 1/50 dilution (1 ug)/Red compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.
Negative control: RAW 264.7(PMID:28060378)

Gated on viable cell.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD73 antibody [RM2050] - BSA and Azide free (ab317463)
This data was developed using Anti-CD73 antibody [RM2050] ab317462, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human stomach tissue labeling CD73 with Anti-CD73 antibody [RM2050] ab317462 at 1/100 (4.97 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Low expression: Weakly positive staining on blood vessels of human stomach. The section was incubated with Anti-CD73 antibody [RM2050] ab317462 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD73 antibody [RM2050] - BSA and Azide free (ab317463)
This data was developed using Anti-CD73 antibody [RM2050] ab317462, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human liver cancer tissue labeling CD73 with Anti-CD73 antibody [RM2050] ab317462 at 1/100 (4.97 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on human liver cancer (PMID: 30971294). The section was incubated with Anti-CD73 antibody [RM2050] ab317462 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD73 antibody [RM2050] - BSA and Azide free (ab317463)
This data was developed using Anti-CD73 antibody [RM2050] ab317462, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling CD73 with Anti-CD73 antibody [RM2050] ab317462 at 1/100 (4.97 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on human spleen. The section was incubated with Anti-CD73 antibody [RM2050] ab317462 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunocytochemistry/ Immunofluorescence - Anti-CD73 antibody [RM2050] - BSA and Azide free (ab317463)
This data was developed using Anti-CD73 antibody [RM2050] ab317462, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 4T1 (mouse mammary gland carcinoma epithelial cell) cells labelling CD73 with Anti-CD73 antibody [RM2050] ab317462 at 1/500 (0.994 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/mL dilution (Green).
Confocal image showing membrane and cytoplasmic staining in 4T1 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).

Negative control: RAW 264.7 (PMID: 28060378)

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.
Immunocytochemistry/ Immunofluorescence - Anti-CD73 antibody [RM2050] - BSA and Azide free (ab317463)
This data was developed using Anti-CD73 antibody [RM2050] ab317462, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NT5E KO A431(human epidermoid carcinoma epithelial cell) cells labelling CD73 with Anti-CD73 antibody [RM2050] ab317462 at 1/500 (0.994 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/mL dilution (Green).
Confocal image showing membrane and cytoplasmic staining in parental A431 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.
Immunoprecipitation - Anti-CD73 antibody [RM2050] - BSA and Azide free (ab317463)
This data was developed using Anti-CD73 antibody [RM2050] ab317462, the same antibody clone in a different buffer formulation.
CD73 was immunoprecipitated from 0.35 mg 4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate with Anti-CD73 antibody [RM2050] ab317462 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-CD73 antibody [RM2050] ab317462 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: 4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate

Lane 2: Anti-CD73 antibody [RM2050] ab317462 IP in 4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate

Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-CD73 antibody [RM2050] ab317462 in 4T1 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-CD73 antibody [RM2050] (Anti-CD73 antibody [RM2050] ab317462) at 1/30 dilution
Lane 1: 4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate
Lane 2: Anti-CD73 antibody [RM2050] ab317462 IP in 4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 180s
Immunoprecipitation - Anti-CD73 antibody [RM2050] - BSA and Azide free (ab317463)
This data was developed using Anti-CD73 antibody [RM2050] ab317462, the same antibody clone in a different buffer formulation.
CD73 was immunoprecipitated from 0.35 mg U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate with Anti-CD73 antibody [RM2050] ab317462 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-CD73 antibody [RM2050] ab317462 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate

Lane 2: Anti-CD73 antibody [RM2050] ab317462 IP in U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate

Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-CD73 antibody [RM2050] ab317462 in U-87 MG whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-CD73 antibody [RM2050] (Anti-CD73 antibody [RM2050] ab317462) at 1/30 dilution
Lane 1: U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate
Lane 2: Anti-CD73 antibody [RM2050] ab317462 IP in U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 32s
Western blot - Anti-CD73 antibody [RM2050] - BSA and Azide free (ab317463)
This data was developed using Anti-CD73 antibody [RM2050] ab317462, the same antibody clone in a different buffer formulation.
Negative control: RAW264.7 (PMID:28060378)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
All lanes: Western blot - Anti-CD73 antibody [RM2050] (Anti-CD73 antibody [RM2050] ab317462) at 1/1000 dilution
Lane 1: 4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate at 40 µg
Lane 2: RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 40 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Observed band size: 75 kDa, 36 kDa
Exposure time: 180s
Western blot - Anti-CD73 antibody [RM2050] - BSA and Azide free (ab317463)
This data was developed using Anti-CD73 antibody [RM2050] ab317462, the same antibody clone in a different buffer formulation.
Negative control: HeLa and Raji (PMID: 8566797)
In Western blot, Anti-CD73 antibody [RM2050] ab317462 was shown to bind specifically to CD73. A band was observed at 75 kDa in wild-type A431 cell lysates (lane 5) whereas loss of signal was observed in the CD73 knockout cell line (lane 6, Human NT5E knockout A-431 cell line ab261895/knockout cell lysate Human NT5E knockout A-431 cell lysate ab261704).
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
All lanes: Western blot - Anti-CD73 antibody [RM2050] (Anti-CD73 antibody [RM2050] ab317462) at 1/1000 dilution
Lane 1: U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 40 µg
Lane 2: A375 (human malignant melanoma epithelial cell) whole cell lysate at 40 µg
Lane 3: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 40 µg
Lane 4: Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate at 40 µg
Lane 5: Wild-type A431 (human epidermoid carcinoma epithelial cell) whole cell lysate at 40 µg
Lane 6: CD73 knockout A431 whole cell lysate at 40 µg
Lane 7: Human spleen tissue lysate at 40 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 75 kDa, 36 kDa
Exposure time: 180s