WB
Western blot analysis of NM23 on different lysates with Mouse anti-NM23 antibody 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), Exposure time: 1 minute; 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 at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody at 1:50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human colon tissue with Mouse anti-NM23 antibody 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 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.IHC
mmunohistochemical analysis of paraffin-embedded mouse kidney tissue with Mouse anti-NM23 antibody 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 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.IHC
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Mouse anti-NM23 antibody 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 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.ICC/IF
Immunocytochemistry analysis of MCF7 cells labeling NM23 with Mouse anti-NM23 antibody 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 at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (594) were used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of MCF7 cells labeling NM23. Cells were fixed and permeabilized. Then stained with the primary antibody (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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Product Name | NM23 Recombinant Mouse Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide within Human NM23 aa 78-126 / 152. |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG1 |
| Host Species | Mouse |
| Tested Applications | FCICC/IFIHCWB |
| WB:1:1000-1:2000 IHC:1:1000 ICC/IF:1:100 FC:1:1000 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | NME1 |
|---|---|
| Gene Synonyms | NB AWD NBS GAAD NDKA NM23 NDPKA NDPK-A NM23-H1 |
| Gene Full Name | NME/NM23 nucleoside diphosphate kinase 1 |
| Gene Summary | This gene (NME1) was identified because of its reduced mRNA transcript levels in highly metastatic cells. Nucleoside diphosphate kinase (NDK) exists as a hexamer composed of 'A' (encoded by this gene) and 'B' (encoded by NME2) isoforms. Mutations in this gene have been identified in aggressive neuroblastomas. Two transcript variants encoding different isoforms have been found for this gene. Co-transcription of this gene and the neighboring downstream gene (NME2) generates naturally-occurring transcripts (NME1-NME2), which encodes a fusion protein comprised of sequence sharing identity with each individual gene product. [provided by RefSeq, Jul 2008] |
| Alternative Names | AWD antibody AWD drosophila homolog of antibody GAAD antibody Granzyme A activated DNase antibody Granzyme A-activated DNase antibody GZMA activated DNase antibody Metastasis inhibition factor NM23 antibody NB antibody NBS antibody NDK A antibody AWD antibody AWD drosophila homolog of antibody GAAD antibody Granzyme A activated DNase antibody Granzyme A-activated DNase antibody GZMA activated DNase antibody Metastasis inhibition factor NM23 antibody NB antibody NBS antibody NDK A antibody NDKA antibody NDKA_HUMAN antibody NDP kinase A antibody NDPK-A antibody NDPKA antibody NM23 antibody NM23 long variant included antibody nm23-H1 antibody NM23-M1 antibody NM23H1B included antibody NME/NM23 nucleoside diphosphate kinase 1 antibody Nme1 antibody NME1-NME2 spliced read-through transcript included antibody Non-metastatic cells 1 protein (NM23A) expressed in antibody Nonmetastatic cells 1 protein expressed in antibody Nonmetastatic protein 23 antibody Nonmetastatic protein 23 homolog 1 antibody Nucleoside diphosphate kinase A antibody Tumor metastatic process-associated protein antibody |
| Molecular Weight(MW) | 17kDa(Observed band size: 17/19kDa) |
| Cellular Localization | Cytoplasm, Nucleus. |
WB
Western blot analysis of NM23 on different lysates with Mouse anti-NM23 antibody 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), Exposure time: 1 minute; 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 at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody at 1:50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human colon tissue with Mouse anti-NM23 antibody 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 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.
IHC
mmunohistochemical analysis of paraffin-embedded mouse kidney tissue with Mouse anti-NM23 antibody 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 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.
IHC
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Mouse anti-NM23 antibody 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 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.
ICC/IF
Immunocytochemistry analysis of MCF7 cells labeling NM23 with Mouse anti-NM23 antibody 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 at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (594) were used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of MCF7 cells labeling NM23. Cells were fixed and permeabilized. Then stained with the primary antibody (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 at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Application Notes | WB:1:1000-1:2000 IHC:1:1000 ICC/IF:1:100 FC:1:1000 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | 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. |
