NM23 Mouse Monoclonal Antibody [12A1]
cat.: EM1801-20
Product Type: Mouse monoclonal IgG1, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, FC
Clonality: Monoclonal
Clone number: 12A1
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 2ug/ul
Purification: Protein G affinity purified.
Molecular weight: Predicted band size: 17 kDa
Isotype: IgG1
Immunogen: Synthetic peptide within Human NM23 aa 78-126 / 152.
Positive control: HeLa cell lysate, A431 cell lysate, A549 cell lysate, MCF7 cell lysate, Caco-2 cell lysate, PC-3M cell lysate, HepG2 cell lysate, NIH/3T3 cell lysate, C6 cell lysate, rat brain tissue lysate, human breast cancer tissue, human liver cancer tissue, human thyroid gland tissue, MCF7.
Subcellular location: Cytoplasm, Nucleus.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
1:1,000-1:2,000
1:50-1:200
1:50-1:200
1:1,000
Uniprot #: SwissProt: P15531 Human | P15532 Mouse | Q05982 Rat
Alternative names: AWD AWD, drosophila, homolog of GAAD Granzyme A activated DNase Granzyme A-activated DNase GZMA activated DNase Metastasis inhibition factor NM23 NB NBS NDK A NDKA NDKA_HUMAN NDP kinase A NDPK-A NDPKA NM23 NM23 long variant, included nm23-H1 NM23-M1 NM23H1B, included NME/NM23 nucleoside diphosphate kinase 1 Nme1 NME1-NME2 spliced read-through transcript, included Non-metastatic cells 1, protein (NM23A) expressed in Nonmetastatic cells 1, protein expressed in Nonmetastatic protein 23 Nonmetastatic protein 23, homolog 1 Nucleoside diphosphate kinase A Tumor metastatic process-associated protein
Images
EM1801-20_1.jpg Fig1: Western blot analysis of NM23 on different lysates with Mouse anti-NM23 antibody (EM1801-20) at 1/2,000 dilution.

Lane 1: HeLa cell lysate (15 µg/Lane)
Lane 2: A431 cell lysate (15 µg/Lane)
Lane 3: A549 cell lysate (15 µg/Lane)
Lane 4: MCF7 cell lysate (15 µg/Lane)
Lane 5: Caco-2 cell lysate (15 µg/Lane)

Predicted band size: 17 kDa
Observed band size: 17/19 kDa

Exposure time: 2 minutes;

4-20% 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 (EM1801-20) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:50,000 dilution was used for 1 hour at room temperature.
EM1801-20_2.jpg Fig2: Western blot analysis of NM23 on different lysates with Mouse anti-NM23 antibody (EM1801-20) at 1/1,000 dilution.

Lane 1: MCF7 cell lysate (20 µg/Lane)
Lane 2: PC-3M cell lysate (20 µg/Lane)
Lane 3: HeLa cell lysate (20 µg/Lane)
Lane 4: HepG2 cell lysate (20 µg/Lane)
Lane 5: A549 cell lysate (20 µg/Lane)
Lane 4: NIH/3T3 cell lysate (20 µg/Lane)
Lane 5: C6 cell lysate (20 µg/Lane)
Lane 5: Rat brain tissue lysate (40 µg/Lane)

Predicted band size: 17 kDa
Observed band size: 17/19 kDa

Exposure time: 3 minutes;

4-20% 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 (EM1801-20) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:50,000 dilution was used for 1 hour at room temperature.
EM1801-20_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human breast cancer tissue using anti-NM23 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody (EM1801-20) at 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
EM1801-20_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human liver cancer tissue using anti-NM23 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody (EM1801-20) at 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
EM1801-20_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human thyroid gland tissue using anti-NM23 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the antibody (EM1801-20) at 1/200 dilution, for 30 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chrogen. Counter stained with hematoxylin and mounted with DPX.
EM1801-20_6.jpg Fig6: Flow cytometric analysis of MCF7 cells labeling NM23.

Cells were fixed and permeabilized. Then stained with the primary antibody (EM1801-20, 1μg/mL) (red) compared with Mouse IgG 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).
EM1801-20_7.jpg Fig7: Immunocytochemistry analysis of MCF7 cells labeling NM23 with Mouse anti-NM23 antibody (EM1801-20) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-NM23 antibody (EM1801-20) at 1/100 dilution in 1% BSA in PBST 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.

beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.