Anti-Niemann Pick C1/NPC1 Antibody Picoband™

Product Name

Anti-Niemann Pick C1/NPC1 Antibody Picoband™

View all Niemann-Pick C1 Antibodies

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.

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.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

A00428-3 is reactive to NPC1 in Human, Mouse, Rat

Applications

A00428-3 is guaranteed for Flow Cytometry, WB Boster Guarantee

Observed Molecular Weight

180 kDa

Calculated molecular weight

142.167kDa

Background of Niemann-Pick C1

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.

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Reconsitution

Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.

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, 1-3 μg/1x10⁶ cells, Human

Validation Images & Assay Conditions

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Figure 1. 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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Figure 2. 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). 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 (Red line) was also used as a control.

Gene/Protein Information For NPC1 (Source: Uniprot.org, NCBI)

Alternative Names

FLJ98532; Niemann-Pick C1 protein; Niemann-Pick disease, type C1; Niemann-Pick Type C1; NPC; NPC1; SLC65A1 NPC1 NPC, POGZ, SLC65A1 NPC intracellular cholesterol transporter 1 NPC intracellular cholesterol transporter 1|Niemann-Pick C1 protein|truncated Niemann-Pick C1

For more info on NPC1, check out the NPC1 Infographic

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In this infographic, you will see the following information for NPC1: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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