You have no items in your shopping cart.
You have no items in your shopping cart.
Catalog Number | orb654447 |
---|---|
Category | Antibodies |
Description | NNT Antibody |
Species/Host | Rabbit |
Clonality | Polyclonal |
Tested applications | ELISA, FC, ICC, IF, WB |
Reactivity | Human, Monkey, Mouse, Rat |
Isotype | Rabbit IgG |
Immunogen | E.coli-derived human NNT recombinant protein (Position: C44-A350). |
Concentration | Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml. |
Dilution range | Western blot, 0.1-0.25μg/ml, Human, Mouse, Monkey, Rat Immunocytochemistry/Immunofluorescence, 4μg/ml, Human Flow Cytometry, 1-3μg/1x106 cells, Human Direct ELISA, 0.1-0.5μg/ml, Human |
Form/Appearance | Lyophilized |
Conjugation | Unconjugated |
MW | 114 kDa |
UniProt ID | Q13423 |
Storage | Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles. |
Note | For research use only |
Application notes | Tested Species: In-house tested species with positive results. Other applications have not been tested. Optimal dilutions should be determined by end users. Add 0.2ml of distilled water will yield a concentration of 500ug/ml. |
Expiration Date | 12 months from date of receipt. |
Western blot analysis of NNT using anti-NNT antibody (orb654447). 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 50ug of sample under reducing conditions. Lane 1: rat liver tissue lysates, Lane 2: rat kidney tissue lysates, Lane 3: rat heart tissue lysates, Lane 4: mouse liver tissue lysates, Lane 5: mouse kidney tissue lysates, Lane 6: mouse heart tissue lysates, Lane 7: human HeLa whole cell lysates. Lane 8: monkey liver tissue lysates. Lane 9: monkey kidney tissue lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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 (Catalog # orb654447) 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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # orb90503) with Tanon 5200 system. A specific band was detected for NNT at approximately 114KD. The expected band size for NNT is at 114KD.
IF analysis of NNT using anti-NNT antibody (orb654447). NNT was detected in immunocytochemical section of U20S cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (orb90553) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 4μg/mL rabbit anti-NNT Antibody (orb654447) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 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.
Flow Cytometry analysis of THP-1 cells using anti-NNT antibody (orb654447). Overlay histogram showing THP-1 cells stained with orb654447 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-NNT Antibody (orb654447, 1μg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
WB | |
Human, Mouse, Porcine, Rat, Zebrafish | |
Rabbit | |
Polyclonal | |
Unconjugated |
Filter by Rating