MA5-15537

NT5E antibody from Invitrogen Antibodies

CALJA, CD73, eN, eNT, NT5

Western blot

Supportive data in Antibodypedia

Immunocytochemistry

Data presented on provider website

Immunohistochemistry

Supportive data in Antibodypedia

Other assay

Supportive data in Antibodypedia

Provider product page for MA5-15537

Antibody Data

Product number

MA5-15537

Provider

Invitrogen Antibodies

Product name

Anti-CD73 Monoclonal Antibody (1D7)

Provider product page

Antibody type

Monoclonal

Antigen

Recombinant protein fragment

Description

MA5-15537 targets NT5E in IHC applications and shows reactivity with Human samples. The MA5-15537 immunogen is purified recombinant fragment of NT5E expressed in E. Coli. MA5-15537 detects NT5E which has a predicted molecular weight of approximately 70kDa.

Reactivity

Human, Mouse, Rat

Host

Mouse

Isotype

IgG

Antibody clone number

1D7

Vial size

100 µL

Concentration

Lot Dependent

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of CD73 was achieved by transfecting A375 cells with CD73 specific siRNAs (Silencer® select Product # s9735, s9736). Western blot analysis (Fig. a) was performed using whole cell extracts from the CD73 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with CD73 Monoclonal Antibody (1D7) (Product # MA5-15537, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/ml, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to CD73.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of A-375 (Lane 1), Hep G2 (Lane 2), HT-29 (Lane 3), SK-OV-3 (Lane 4), U-2 OS (Lane 5), tissue extracts of Mouse Uterus (Lane 6), Rat Uterus (Lane 7), Mouse Placenta (Lane 8), Mouse Ovary (Lane 9) and Mouse Lung (Lane 10). The blot was probed with CD73 Monoclonal Antibody (Product # MA5-15537, 1:500 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/ml, 1:4000 dilution). A band at 63 kDa corresponding to CD73 was observed across the cell lines and tissues tested.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of paraffin-embedded human lung cancer (A), cholangiocarcinoma (B), lymph node (C) and esophagus (D) tissues using CD73/NT5E monoclonal antibody (Product # MA5-15537) followed with DAB staining.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of paraffin-embedded human lung cancer (A), cholangiocarcinoma (B), lymph node (C) and esophagus (D) tissues using CD73/NT5E monoclonal antibody (Product # MA5-15537) followed with DAB staining.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of CD73 was performed using 70% confluent log phase A-375 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with CD73 Monoclonal Antibody (1D7) (Product # MA5-15537) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415). Panel d represents the merged image showing membranous and cytosolic localization. The images were captured at 60X magnification.