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Validated using a knockout cell line

Anti-CD73 antibody [4G6E3] (ab202122)

Reviews (1) Submit a question References (10)
Western blot - Anti-CD73 antibody [4G6E3] (ab202122)
  • Flow Cytometry - Anti-CD73 antibody [4G6E3] (ab202122)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD73 antibody [4G6E3] (ab202122)
  • Western blot - Anti-CD73 antibody [4G6E3] (ab202122)
  • Western blot - Anti-CD73 antibody [4G6E3] (ab202122)
  • Western blot - Anti-CD73 antibody [4G6E3] (ab202122)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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

  • 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

  • Immunogen

    Recombinant fragment corresponding to Human CD73 aa 1-250. Expressed in E. Coli.
    Database link: P21589

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • Positive control

    • Flow Cytometry: A431 cells; WB: A375, and A431 cell lysates; IHC-P: Human endometrial cancer and Human esophageal cancer tissues.

  • 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

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.
WB
1/500 - 1/2000.

Target

  • 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

  • Database links

    see all

  • Alternative names

    • 5' NT antibody
    • 5' nucleotidase (CD73) antibody
    • 5' nucleotidase precursor antibody
    • 5' nucleotidase, ecto antibody
    • 5' nucleotidase, ecto (CD73) antibody
    • 5'-NT antibody
    • 5'-nucleotidase antibody
    • 5NTD_HUMAN antibody
    • CD73 antibody
    • CD73 antigen antibody
    • E5NT antibody
    • Ecto 5' nucleotidase antibody
    • Ecto-5'-nucleotidase antibody
    • eN antibody
    • eNT antibody
    • NT antibody
    • NT5 antibody
    • NT5E antibody
    • NTE antibody
    • Purine 5 Prime Nucleotidase antibody
    see all

Images

  • Western blot - Anti-CD73 antibody [4G6E3] (ab202122)
    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.

  • Flow Cytometry - Anti-CD73 antibody [4G6E3] (ab202122)
    Flow Cytometry - Anti-CD73 antibody [4G6E3] (ab202122)

    Flow cytometric analysis of A431 cells using ab202122 at 1/200 (green) and negative control (red).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD73 antibody [4G6E3] (ab202122)
    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.

  • Western blot - Anti-CD73 antibody [4G6E3] (ab202122)
    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.

  • Western blot - Anti-CD73 antibody [4G6E3] (ab202122)
    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)
    Western blot - Anti-CD73 antibody [4G6E3] (ab202122)
    Anti-CD73 antibody [4G6E3] (ab202122) at 1/500 dilution + Human CD73 (AA: 30-250) recombinant protein


    Expected MW is 26.6 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD73 antibody [4G6E3] (ab202122)
    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.

References (10)

Publishing research using ab202122? Please let us know so that we can cite the reference in this datasheet.

ab202122 has been referenced in 10 publications.

  • Li X LncRNA metastasis-associated lung adenocarcinoma transcript-1 promotes osteogenic differentiation of bone marrow stem cells and inhibits osteoclastic differentiation of Mø in osteoporosis via the miR-124-3p/IGF2BP1/Wnt/β-catenin axis. J Tissue Eng Regen Med 16:311-329 (2022). PubMed: 34962086
  • Xie A  et al. Kartogenin Induced Adipose-Derived Stem Cell Exosomes Enhance the Chondrogenic Differentiation Ability of Adipose-Derived Stem Cells. Dis Markers 2022:6943630 (2022). PubMed: 36072901
  • Zheng WV  et al. Anti-aging effect of β-carotene through regulating the KAT7-P15 signaling axis, inflammation and oxidative stress process. Cell Mol Biol Lett 27:86 (2022). PubMed: 36209059
  • Hu M  et al. The function and mechanism of the miR-210-3p/KRAS axis in bone marrow-derived mesenchymal stem cell from patients with osteoporosis. J Tissue Eng Regen Med 15:699-711 (2021). PubMed: 33982450
  • Ren X  et al. Osteogenic ability using porous hydroxyapatite scaffold-based delivery of human placenta-derived mesenchymal stem cells. Exp Ther Med 22:1091 (2021). PubMed: 34504545
  • Zhang J  et al. A novel circular RNA circ_HN1/miR-628-5p/Ecto-5'-nucleotidase competing endogenous RNA network regulates gastric cancer development. Bioengineered 12:9739-9752 (2021). PubMed: 34637682
  • 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
  • Baracho Trindade Hill A  et al. Placental scaffolds have the ability to support adipose-derived cells differentiation into osteogenic and chondrogenic lineages. J Tissue Eng Regen Med 14:1661-1672 (2020). PubMed: 32893450
  • 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

Customer reviews and Q&As

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD73 antibody [4G6E3]

 Good
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Sheep Tissue sections (Kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Sodium Citrate Buffer pH6.0
Permeabilization
No
Specification
Kidney
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Verified customer

Submitted Nov 28 2016

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