NNT Rabbit Polyclonal Antibody

SKU: orb654447

Description

NNT Rabbit Polyclonal Antibody

Research Area

Cancer Biology, Metabolism Research, Signal Transduction

Images & Validation

• Tested Applications: ELISA, FC, ICC, IF, WB
• Dilution Range: Western blot, 0.1-0.25μg/ml, Human, Mouse, Monkey, Rat, Pig Immunocytochemistry/Immunofluorescence, 5μg/ml, Human Flow Cytometry (Fixed), 1-3μg/1x10^6 cells, Human ELISA, 0.1-0.5μg/ml, -
• Reactivity: Human, Monkey, Mouse, Porcine, Rat

Key Properties

• Antibody Type: Primary Antibody
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Immunogen: E.coli-derived human NNT recombinant protein (Position: C44-A350).
• Target: NAD
• Molecular Weight: 114 kDa
• Purification: Immunogen affinity purified.
• Conjugation: Unconjugated

Storage & Handling

• 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.
• Form/Appearance: Lyophilized
• Buffer/Preservatives: Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
• Concentration: 500 µg/ml
• Expiration Date: 12 months from date of receipt.
• Disclaimer: For research use only

Alternative Names

NNT

Flow Cytometry analysis of U937 cells using anti-NNT antibody. Overlay histogram showing U937 cells (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-NNT Antibody (1 µg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10 µg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

IF analysis of NNT using anti-NNT antibody. NNT was detected in immunocytochemical section of SiHa cells. 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 5 µg/mL rabbit anti-NNT Antibody overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of NNT using anti-NNT antibody. Electrophoresis was performed on a 8% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: human Hela whole cell lysates, Lane 2: human SiHa whole cell lysates, Lane 3: monkey kidney tissue lysates, Lane 4: pig kidney tissue lysates, Lane 5: rat kidney tissue lysates, Lane 6: 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-NNT antigen affinity purified polyclonal antibody at 0.25 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 ECL Plus Western Blotting Substrate with Tanon 5200 system. A specific band was detected for NNT at approximately 114 kDa. The expected band size for NNT is at 114 kDa.

Quick Database Links

Gene Symbol

NAD

UniProt

Q13423