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Product Info Summary
| SKU: | A00428-3 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | Flow Cytometry, WB |
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Product info
Product Name
Anti-Niemann Pick C1/NPC1 Antibody Picoband®
SKU/Catalog Number
A00428-3
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Niemann Pick C1/NPC1 Antibody Picoband® catalog # A00428-3. Tested in Flow Cytometry, WB applications. This antibody reacts with Human, Mouse, Rat. 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
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 freezing and thawing.
Cite This Product
Anti-Niemann Pick C1/NPC1 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00428-3)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
A synthetic peptide corresponding to a sequence in the middle region of human Niemann Pick C1/NPC1.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00428-3 is reactive to NPC1 in Human, Mouse, Rat
Observed Molecular Weight
180 kDa
Calculated molecular weight
142.2 kDa
Background of NPC1
Niemann-Pick disease, type C1 (NPC1) is a membrane protein that mediates intracellular cholesterol trafficking in mammals. In humans it is encoded by the NPC1 gene (chromosome location 18q11). This gene encodes a large protein that resides in the limiting membrane of endosomes and lysosomes and mediates intracellular cholesterol trafficking via binding of cholesterol to its N-terminal domain. It is predicted to have a cytoplasmic C-terminus, 13 transmembrane domains, and 3 large loops in the lumen of the endosome - the last loop being at the N-terminus. This protein transports low-absorbance lipoproteins to late endosomal/lysosomal compartments where they are hydrolized and released as free cholesterol. Defects in this gene cause Niemann-Pick type C disease, a rare autosomal recessive neurodegenerative disorder characterized by over accumulation of cholesterol and glycosphingolipids in late endosomal/lysosomal compartments.
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
A00428-3 is guaranteed for Flow Cytometry, 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, 0.25-0.5 μg/ml, Human, Mouse, Rat
Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human
Positive Control
WB: human 293T whole cell, human HepG2 whole cell, human Hela whole cell, rat PC-12 whole cell
FCM: HepG2 cell
Validation Images & Assay Conditions
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Western blot analysis of Niemann Pick C1/NPC1 using anti-Niemann Pick C1/NPC1 antibody (A00428-3).
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 293T whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human Hela whole cell lysates,
Lane 4: rat PC-12 whole cell 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-Niemann Pick C1/NPC1 antigen affinity purified polyclonal antibody (Catalog # A00428-3) 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 (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Niemann Pick C1/NPC1 at approximately 180 kDa. The expected band size for Niemann Pick C1/NPC1 is at 142 kDa.
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Flow Cytometry analysis of HepG2 cells using anti-Niemann Pick C1/NPC1 antibody (A00428-3).
Overlay histogram showing HepG2 cells stained with A00428-3 (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-Niemann Pick C1/NPC1 Antibody (A00428-3, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 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.
Specific Publications For Anti-Niemann Pick C1/NPC1 Antibody Picoband® (A00428-3)
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