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Product Info Summary
| SKU: | A00887-3 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Monkey, Mouse, Pig, Rat |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-NNT Antibody Picoband®
SKU/Catalog Number
A00887-3
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-NNT Antibody Picoband® catalog # A00887-3. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Monkey, Mouse, Rat, Pig. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
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.
Cite This Product
Anti-NNT Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00887-3)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human NNT recombinant protein (Position: C44-A350).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00887-3 is reactive to NNT in Human, Monkey, Mouse, Pig, Rat
Observed Molecular Weight
114 kDa
Calculated molecular weight
113.9 kDa
Background of NNT
This gene encodes an integral protein of the inner mitochondrial membrane. The enzyme couples hydride transfer between NAD (H) and NADP (+) to proton translocation across the inner mitochondrial membrane. Under most physiological conditions, the enzyme uses energy from the mitochondrial proton gradient to produce high concentrations of NADPH. The resulting NADPH is used for biosynthesis and in free radical detoxification.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
A00887-3 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.1-0.25μg/ml, Human, Mouse, Monkey, Rat, Pig
Immunocytochemistry/Immunofluorescence, 5μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
ELISA, 0.1-0.5μg/ml, -
Positive Control
WB: human Hela whole cell, human SiHa whole cell, monkey kidney tissue, pig kidney tissue, rat kidney tissue, mouse kidney tissue
ICC/IF: SiHa cell
FCM: U937 cell
Validation Images & Assay Conditions
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Western blot analysis of NNT using anti-NNT antibody (A00887-3).
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 (A00887-3) 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 (Catalog # BA1054) at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) 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.
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IF analysis of NNT using anti-NNT antibody (A00887-3).
NNT was detected in immunocytochemical section of SiHa cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-NNT Antibody (A00887-3) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) 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.
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Flow Cytometry analysis of U937 cells using anti-NNT antibody (A00887-3).
Overlay histogram showing U937 cells stained with A00887-3 (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 (A00887-3,1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Specific Publications For Anti-NNT Antibody Picoband® (A00887-3)
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