CD73 Mouse Monoclonal Antibody [A6F7]
cat.: HA601010
Product Type: Mouse monoclonal IgG1, primary antibodies
Species reactivity: Human
Applications: WB, IHC-P, IF-Tissue, FC
Clonality: Monoclonal
Clone number: A6F7
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein G affinity purified.
Molecular weight: Predicted band size: 63 kDa
Isotype: IgG1
Immunogen: Recombinant protein within Human CD73 27-549.
Positive control: U87MG cell lysates, NCI-H441 xenograft tissue, U87MG xenograft tissue, U87-MG.
Subcellular location: Cell membrane.
Recommended Dilutions:
  WB
  IHC-P
  IF-Tissue
  FC

1:500
1:200-1:600
1:100
1:500-1:1,000
Uniprot #: SwissProt: P21589 Human
Alternative names: 5' NT 5' nucleotidase (CD73) 5' nucleotidase precursor 5' nucleotidase, ecto 5' nucleotidase, ecto (CD73) 5'-NT 5'-nucleotidase 5NTD_HUMAN CD73 CD73 antigen E5NT Ecto 5' nucleotidase Ecto-5'-nucleotidase eN eNT NT NT5 NT5E NTE Purine 5 Prime Nucleotidase
Images
HA601010_1.jpg Fig1: Western blot analysis of CD73 on U87MG cell lysates with Mouse anti-CD73 antibody (HA601010) at 1/500 dilution.

Lysates/proteins at 10 µg/Lane.

Predicted band size: 63 kDa
Observed band size: 70 kDa

Exposure time: 15 seconds;

12% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA601010) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:100,000 dilution was used for 1 hour at room temperature.
HA601010_2.jpg Fig2: Immunohistochemical analysis of paraffin-embedded NCI-H441 xenograft tissue with Mouse anti-CD73 antibody (HA601010) at 1/600 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA601010) at 1/600 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA601010_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded U87MG xenograft tissue with Mouse anti-CD73 antibody (HA601010) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA601010) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA601010_4.jpg Fig4: Immunocytochemistry analysis of U87-MG cells labeling CD73 with Mouse anti-CD73 antibody (HA601010) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, blocked with 2% BSA for 30 minutes at room temperature. Cells were then incubated with Mouse anti-CD73 antibody (HA601010) at 1/100 dilution in 2% BSA overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
HA601010_5.jpg Fig5: Flow cytometric analysis of U87-MG cells labeling CD73.

Cells were fixed and permeabilized.Then stained with the primary antibody (HA601010, 1ug/ml) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.