Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free
- RabMAb
- Recombinant
- KO Validated
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(1 Publication)
Rabbit Recombinant Monoclonal Niemann Pick C2 antibody. Carrier free. Suitable for WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Mouse, Rat, Human samples. Cited in 1 publication.
View Alternative Names
HE1, NPC2, NPC intracellular cholesterol transporter 2, Epididymal secretory protein E1, Human epididymis-specific protein 1, Niemann-Pick disease type C2 protein, He1
- IHC-P
Lab
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (AB251560)
This data was developed using ab218192, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human breast carcinoma labelling Niemann Pick C2 with ab218192 at 1/8000 dilution, followed by a Goat Anti-Rabbit IgG H&L (HRP polymer) ready to use (ab214880). Positive staining on Human breast carcinoma is observed. Counter stained with hematoxylin. Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a Goat Anti-Rabbit IgG H&L (HRP polymer) ready to use. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
- Flow Cyt (Intra)
Supplier Data
Flow Cytometry (Intracellular) - Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (AB251560)
This data was developed using ab218192, the same antibody clone in a different buffer formulation.
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling Niemann Pick C2 with ab218192 at 1/70 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
- IHC-P
Supplier Data
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (AB251560)
This data was developed using ab218192, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human testis tissue labeling Niemann Pick C2 with ab218192 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Granularly cytoplasmic staining on human testis [PMID : 8924505]. Counter stained with Hematoxylin.
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
- ICC/IF
Supplier Data
Immunocytochemistry/ Immunofluorescence - Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (AB251560)
This data was developed using ab218192, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling Niemann Pick C2 with ab218192 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HepG2 cell line.
The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
- IHC-P
Supplier Data
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (AB251560)
This data was developed using ab218192, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human epididymis tissue labeling Niemann Pick C2 with ab218192 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on human epididymis [PMID : 8924505]. Counter stained with Hematoxylin.
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
- IHC-P
Supplier Data
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (AB251560)
This data was developed using ab218192, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling Niemann Pick C2 with ab218192 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Granularly cytoplasmic staining on rat kidney tubules [PMID : 24147030]. Counter stained with Hematoxylin.
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
- IHC-P
Supplier Data
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (AB251560)
This data was developed using ab218192, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling Niemann Pick C2 with ab218192 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Granularly cytoplasmic staining on mouse kidney tubules [PMID : 24147030]. Counter stained with Hematoxylin.
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
- Flow Cyt (Intra)
Supplier Data
Flow Cytometry (Intracellular) - Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (AB251560)
This data was developed using ab218192, the same antibody clone in a different buffer formulation.
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling Niemann Pick C2 with ab218192 at 1/70 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
- ICC/IF
Supplier Data
Immunocytochemistry/ Immunofluorescence - Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (AB251560)
This data was developed using ab218192, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling Niemann Pick C2 with ab218192 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on NIH/3T3 cell line.
The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
- WB
Supplier Data
Western blot - Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (AB251560)
This data was developed using ab218192, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer : 5% NFDM/TBST.
Exposure times : Lanes 1-4 : 4 seconds; Lanes 5-6 : 15 seconds.
Niemann Pick C2 can be glycosylated, the WB pattern is consistent with references PMID : 16374838; 22183894.
All lanes:
Western blot - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (<a href='/en-us/products/primary-antibodies/niemann-pick-c2-antibody-epr19993-145-1-ab218192'>ab218192</a>) at 1/5000 dilution
Lane 1:
Human fetal kidney lysate at 20 µg
Lane 2:
Mouse kidney lysate at 20 µg
Lane 3:
Rat kidney lysate at 20 µg
Lane 4:
HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg
Lane 5:
Rat spleen lysate at 20 µg
Lane 6:
Human spleen lysate at 20 µg
Secondary
All lanes:
Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution
Predicted band size: 17 kDa
Observed band size: 16-18 kDa
false
- WB
Lab
Western blot - Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (AB251560)
This data was developed using ab218192, the same antibody clone in a different buffer formulation.
Lanes 1-3 : Merged signal (red and green). Green - ab218192 observed at 16-18 kDa. Red - loading control ab8245 observed at 36 kDa.
ab218192 Anti-Niemann Pick C2 antibody [EPR19993-145-1] was shown to specifically react with Niemann Pick C2 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266749 (knockout cell lysate ab258079) was used. Wild-type and Niemann Pick C2 knockout samples were subjected to SDS-PAGE. ab218192 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (<a href='/en-us/products/primary-antibodies/niemann-pick-c2-antibody-epr19993-145-1-ab218192'>ab218192</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2:
NPC2 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2:
Western blot - Human NPC2 (Niemann Pick C2) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-npc2-niemann-pick-c2-knockout-hek-293t-cell-line-ab266749'>ab266749</a>)
Lane 3:
HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 17 kDa
Observed band size: 16-18 kDa
false
- WB
Supplier Data
Western blot - Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free (AB251560)
This data was developed using ab218192, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer : 5% NFDM/TBST.
Niemann Pick C2 can be glycosylated, the WB pattern is consistent with references PMID : 16374838; 22183894.
All lanes:
Western blot - Anti-Niemann Pick C2 antibody [EPR19993-145-1] (<a href='/en-us/products/primary-antibodies/niemann-pick-c2-antibody-epr19993-145-1-ab218192'>ab218192</a>) at 1/2000 dilution
Lane 1:
C6 (Rat glial tumor cell line) whole cell lysate at 10 µg
Lane 2:
RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg
Lane 3:
PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg
Lane 4:
NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 10 µg
Secondary
All lanes:
Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution
Predicted band size: 17 kDa
Observed band size: 16-18 kDa
false
Exposure time: 10s
Related conjugates and formulations (1)
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Anti-Niemann Pick C2 antibody [EPR19993-145-1]
Reactivity data
Product details
ab251560 is the carrier-free version of ab218192.
Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free batch production
For more information, read more on recombinant antibodies.
Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
Compatibility
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
Properties and storage information
Form
Purification technique
Storage buffer
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Storage information
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
NPC2 protein facilitates the transfer of cholesterol from lysosomes to other parts of the cell. It operates in tandem with its partner protein Niemann Pick C1 (NPC1) to mediate the egress of cholesterol. The NPC2 protein docks to NPC1 within the lysosomal membrane forming an essential complex in lipid transport. This function is pivotal for the maintenance of cellular cholesterol homeostasis influencing various cellular and systemic functions linked to cholesterol metabolism.
Pathways
NPC2 protein participates prominently in the cholesterol efflux pathways. It associates with the NPC1 protein to regulate the movement of cholesterol from the lysosomal membrane to cytoplasmic areas where it can be utilized or stored. This pathway links NPC2 to significant biological processes like the homeostasis of lipoprotein-derived cholesterol influencing the SREBP pathway which governs lipid synthesis and uptake. Such interactions highlight NPC2's involvement in regulating cellular lipid balance.
Product protocols
- Visit the General protocols
- Visit the Troubleshooting
Target data
Publications (1)
Recent publications for all applications. Explore the full list and refine your search
Cancer cell 39:1479-1496.e18 PubMed34653364
2021
Applications
Unspecified application
Species
Unspecified reactive species
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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