NM23A/NME1 Antibody

• Catalog Number: orb76342
• Category: Antibodies
• Description: NM23A/NME1 Antibody
• Clonality: Polyclonal
• Species/Host: Rabbit
• Isotype: Rabbit IgG
• Conjugation: Unconjugated
• Reactivity: Human, Mouse, Rat
• Form/Appearance: Lyophilized
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Purification: Immunogen affinity purified.
• Immunogen: A synthetic peptide corresponding to a sequence at the C-terminus of human NM23A, different from the related mouse sequence by two amino acids and from rat sequence by one amino acid.
• UniProt ID: P15531
• MW: 17 kDa
• Tested applications: FC, ICC, IHC, IHC-Fr, WB
• Application notes: Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, RatWestern blot, 0.1-0.5μg/ml, Human, Rat, MouseImmunohistochemistry (Frozen Section), 0.5-1μg/ml, Human Immunocytochemistry, 0.5-1μg/ml, Human Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
• Cross Reactivity: No cross-reactivity with other proteins
• Antibody Type: Primary Antibody
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Alternative names: Nucleoside diphosphate kinase A; NDK A; NDP kinase
• Note: For research use only

Western blot analysis of NM23A using anti-NM23A antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: human A549 whole cell lysates, Lane 2: human MCF-7 whole cell lysates, Lane 3: human Hela whole cell lysates, Lane 4: human PC-3 whole cell lysates, Lane 5: rat brain tissue lysates, Lane 6: rat kidney tissue lysates, Lane 7: mouse brain tissue lysates, Lane 8: mouse kidney tissue lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-NM23A antigen affinity purified polyclonal antibody at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for NM23A at approximately 17 kDa. The expected band size for NM23A is at 17 kDa.

IHC analysis of NM23A using anti-NM23A antibody. NM23A was detected in paraffin-embedded section of Rat Cerebellum tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-NM23A Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

IHC analysis of NM23A using anti-NM23A antibody. NM23A was detected in immunocytochemical section of HELA Cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 1 μg/ml rabbit anti-NM23A Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Flow Cytometry analysis of Hela cells using anti-NME1 antibody. Overlay histogram showing Hela cells stained with orb76342 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-NME1 Antibody (orb76342, 1 μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10^6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

IHC analysis of NM23A using anti-NM23A antibody. NM23A was detected in paraffin-embedded section of human mammary cancer tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-NM23A Antibody overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.