EM1801-20

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, MCF7, human breast cancer tissue, human lymphoma tissue, mouse kidney tissue, rat kidney tissue.
Subcellular location:	Cytoplasm, Nucleus.
Recommended Dilutions: WB IF-Cell IHC-P FC	1:1,000-1:2,000 1:50-1:200 1:200-1:1,000 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

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.

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.

	Fig3: 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Mouse anti-NM23 antibody (EM1801-20) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (EM1801-20) 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded human lymphoma tissue with Mouse anti-NM23 antibody (EM1801-20) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (EM1801-20) at 1/1,000 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.

	Fig6: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Mouse anti-NM23 antibody (EM1801-20) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (EM1801-20) at 1/1,000 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.
	Fig7: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Mouse anti-NM23 antibody (EM1801-20) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (EM1801-20) at 1/1,000 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.
	Fig8: 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).

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