Anti-CD73 (NT5E) Antibody, clone 1F22 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1035 - recombinant, expressed in HEK 293 cells
Synonym: 5′-NT; 5′-Nucleotidase; EC:3.1.3.5; Ecto-5′-nucleotidase
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human tonsil (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:1,000 dilution of Cat. No. ZRB1035, Anti-CD73 (NT5E), clone 1F22 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in both germinal center and non-germinal center cells of human tonsil tissue.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human tonsil (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:1,000 dilution of Cat. No. ZRB1035, Anti-CD73 (NT5E), clone 1F22 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in both germinal center and non-germinal center cells of human tonsil tissue.
Immunohistochemistry (Paraffin) Enhanced Validation-Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) human tonsil (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:1,000 dilution of Cat. No. ZRB1035, Anti-CD73 (NT5E), clone 1F22 ZooMAb® Rabbit Monoclonal. Reactivity was detected using an Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in both germinal center and non-germinal center cells of human tonsil tissue.
Western Blotting Enhanced Validation-Recombinant Antibody Technology Lysates from HT-29 cells (Lane 1), A375 cells(Lane 2), Rat Spleen tissue (Lane 3), and Mouse Spleen tissue (Lane 4) were probed with Cat. No. ZRB1035, Anti-CD73 (NT5E), clone 1F22 ZooMAb® Rabbit Monoclonal (1:1,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates CD73 (NT5E) (~63 kDa).
Affinity Binding Assay Enhanced Validation-Recombinant Antibody Technology 100 mg/mL of non-specific GST protein (Run 1) or recombinant NT5E/CD73 protein (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1035, Anti-CD73 (NT5E), clone 1F22 ZooMAb® Rabbit Monoclonal. The binding of this antibody to recombinant NT5E/CD73 protein displayed a KD value of 4.2 x 10-8 whereas its binding to non-specific GST protein displayed a KD of 1.6 x 10-3.
Flow Cytometry Enhanced Validation-Recombinant Antibody Technology Staining of one million HT29 cells was performed using 0.1 μg of Cat. No. ZRB1035, Anti-CD73 (NT5E), clone 1F22 ZooMAb® Rabbit Monoclonal (Yellow histogram) or the equivalent amount of a Rabbit IgG isotype control (Grey histogram), followed by a PE-conjugated Donkey Anti-RabbitIgG secondary antibody.
12 Principles of Green Chemistry: Principle 1—Waste Prevention. This product has a smaller environmental footprint through reduction of waste in the manufacturing process.
12 Principles of Green Chemistry: Principle 4—Designing Safer Chemicals
Chemical products should be designed to affect their desired function while minimizing their toxicity.
Chemical products should be designed to affect their desired function while minimizing their toxicity.
12 Principles of Green Chemistry: Principle 6—Design for Energy Efficiency. This product was re-engineered to utilize less energy in the manufacturing process.
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| biological source | rabbit |
| clone | 1F22, monoclonal |
| recombinant monoclonal | |
| conjugate | unconjugated |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| form | lyophilized |
| greener alternative category | Aligned |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry . |
| isotype | IgG |
| mol wt | calculated mol wt 63.37 kDa |
| observed mol wt ~63 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | mouse, rat, human |
| storage temp. | 2-8°C |
| technique(s) | affinity binding assay: suitable |
| flow cytometry: suitable | |
| immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable | |
| western blot: suitable | |
| UniProt accession no. | P21589  |
| Application: | Anti-CD73 (NT5E), clone 1F22 ZooMAb, Cat. No. ZRB1035, is a recombinant Rabbit monoclonal antibody that detects CD73 (NT5E) and is tested for use in Affinity Binding Assay, Flow Cytometry, Immunohistochemistry (Paraffin), and Western Blotting. |
| Application: | Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected CD73 (NT5E) in A375 cells and in rat spleen and mouse spleen tissue lysates. Affinity Binding Assay: A representative lot of this antibody bound recombinant NT5E/CD73 protein with a KD of 4.2 x 10-8 in an affinity binding assay. Immunohistochemistry (Paraffin) Analysis: A 1:1,000 dilution from a representative lot detected CD73 (NT5E) in human tonsil tissue sections. Flow Cytometry Analysis: 0.1 μg from a representative lot detected CD73 (NT5E) in one million HT29 cells. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| Disclaimer: | Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals. |
| General description: | We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information. |
| General description: | ZooMAb antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb antibodies are reliably available and ready to ship when you need them. Learn more about ZooMAb here. |
| Immunogen: | His-tagged recombinant fragment corresponding to 244 amino acids from the N-terminal half of mature human CD73. |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL. |
| Reconstitution: | 300 μg/mL after reconstitution at 25 μL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type. |
| Specificity: | Clone 1F22 is a ZooMAb rabbit recombinant monoclonal antibody that specifically detects 5′-Nucleotidase (CD73). It targets an epitope within the N-terminal half. |
| Storage and Stability: | Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws. |
| Target description: | 5′-Nucleotidase (UniProt: P21589; also known as EC:3.1.3.5, 5′-NT, Ecto-5′-nucleotidase, CD73) is encoded by the NT5E (also known as NT5, NTE) gene (Gene ID: 4907) in human. CD73 is a GPI-anchor membrane glycoprotein that is expressed in most tissues and is involved in the control of a variety of physiological responses including epithelial ion and fluid transport, ischemic preconditioning, tissue injury, platelet function, hypoxia and vascular leak. It is also widely expressed on several tumor tissues and its higher expression levels are associated with tumor neovascularization, invasiveness, and metastasis. Its biological actions are mainly due to its regulated enzymatic phosphohydrolytic activity on extracellular nucleotides. It hydrolyzes extracellular nucleotides into membrane permeable nucleosides and displays AMP-, NAD-, and NMN-nucleosidase activities. CD73 is synthesized with a signal peptide (aa 1-26) and a C-terminal propeptide (aa 550-574) that are subsequently cleaved off to generate the mature form (aa 27-549). Its substrate binding site is localized to amino acids 500-506. Mutations in NT5E gene have been linked to adult-onset of calcification of joints and arteries. This ZooMAb recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Spychala, J. (2000). Pharmacol. Ther. 87 (2-3); 161-173; Zhang, B. (2010). Cancer Res. 70(16); 6407-6411). |
| WGK Germany | WGK 1 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |