Anti-Niemann Pick C2/NPC2 Picoband™ Antibody

Product Name

Anti-Niemann Pick C2/NPC2 Picoband™ Antibody

View all Niemann-Pick type C2 Antibodies

SKU/Catalog Number

A01582-3

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-Niemann Pick C2/NPC2 Picoband™ Antibody catalog # A01582-3. Tested in ELISA, Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse.

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-Niemann Pick C2/NPC2 Picoband™ Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # A01582-3)

Host

Rabbit

Contents

Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na₂HPO₄, 0.05mg NaN₃.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived human Niemann Pick C2/NPC2 recombinant protein (Position: E20-L151).

*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

A01582-3 is reactive to NPC2 in Human, Mouse

Applications

A01582-3 is guaranteed for ELISA, Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee

Observed Molecular Weight

19 kDa

Calculated molecular weight

16.57kDa

Background of Niemann-Pick type C2

Epididymal secretory protein E1 (also called NPC intracellular cholesterol transporter 2, or NPC2) is a protein associated with Niemann-Pick disease, type C. It is mapped to 14q24.3. This gene encodes a protein containing a lipid recognition domain. The encoded protein may function in regulating the transport of cholesterol through the late endosomal/lysosomal system. Mutations in this gene have been associated with Niemann-Pick disease, type C2 and frontal lobe atrophy.

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

Add 0.2ml of distilled water will yield a concentration of 500ug/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, 0.25-0.5μg/ml, Human
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Flow Cytometry, 1-3μg/1x10⁶ cells, Human
Direct ELISA, 0.1-0.5μg/ml, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of NPC2 using anti-NPC2 antibody (A01582-3).
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 U20S 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-NPC2 antigen affinity purified polyclonal antibody (Catalog # A01582-3) 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 NPC2 at approximately 19 kDa. The expected band size for NPC2 is at 16 kDa.

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Figure 2. IHC analysis of NPC2 using anti-NPC2 antibody (A01582-3).
NPC2 was detected in paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-NPC2 Antibody (A01582-3) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

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Figure 3. IHC analysis of NPC2 using anti-NPC2 antibody (A01582-3).
NPC2 was detected in paraffin-embedded section of mouse spleen tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-NPC2 Antibody (A01582-3) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

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Figure 4. Flow Cytometry analysis of HepG2 cells using anti-NPC2 antibody (A01582-3).
Overlay histogram showing HepG2 cells stained with A01582-3 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-NPC2 Antibody (A01582-3, 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 (Red line) was also used as a control.

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Figure 5. IF analysis of NPC2 using anti-NPC2 antibody (A01582-3).
NPC2 was detected in immunocytochemical section of A431 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 2μg/mL rabbit anti-NPC2 Antibody (A01582-3) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) 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.

Gene/Protein Information For NPC2 (Source: Uniprot.org, NCBI)

Alternative Names

EDDM1; Epididymal Protein 1; epididymal secretory protein E1; HE1; HE1epididymal protein 1; Human epididymis-specific protein 1; MGC1333; Niemann-Pick disease type C2 protein; Niemann-Pick disease, type C2; NiemannPick Type C2; Niemann-Pick Type C2; NPC2; NP-C2; tissue-specific secretory protein NPC2 EDDM1, HE1 NPC intracellular cholesterol transporter 2 NPC intracellular cholesterol transporter 2|Niemann-Pick disease type C2 protein|epididymal protein 1|epididymis secretory sperm binding protein|human epididymis-specific protein 1|tissue-specific secretory protein

For more info on NPC2, check out the NPC2 Infographic

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In this infographic, you will see the following information for NPC2: 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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