重组Anti-Niemann Pick C1抗体[EPR5209] (ab134113)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR5209] to Niemann Pick C1
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-Niemann Pick C1抗体[EPR5209]
参阅全部 Niemann Pick C1 一抗 -
描述
兔单克隆抗体[EPR5209] to Niemann Pick C1 -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide within Human Niemann Pick C1 aa 1250 to the C-terminus (C terminal). The exact sequence is proprietary.
Database link: O15118 -
阳性对照
- WB: Wild-type HAP1 whole cell lysate; HEK-293, HepG2, THP-1, PC-3, 3T3L1, L6 and HEK-293T cell lysates; Rat liver and brain lysates. IHC-P: Human kidney and liver tissues; Mouse liver tissue; Rat cerebellum tissue. ICC/IF: Neuro-2a cells; SH-SY5Y control and PADK treated cells. Flow Cyt (intra): Neuro-2a cells.
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常规说明
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
解离常数(KD)
KD = 4.90 x 10 -11 M Learn more about KD -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR5209 -
同种型
IgG -
研究领域
- Metabolism
- Pathways and Processes
- Metabolic signaling pathways
- Lipid and lipoprotein metabolism
- Cholesterol Metabolism
- Metabolism
- Pathways and Processes
- Metabolic signaling pathways
- Lipid and lipoprotein metabolism
- Lipoprotein metabolism
相关产品
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Alternative Versions
- APC Anti-Niemann Pick C1 antibody [EPR5209] (ab223985)
- Anti-Niemann Pick C1 antibody [EPR5209] - BSA and Azide free (ab224268)
- PE Anti-Niemann Pick C1 antibody [EPR5209] (ab317903)
- Alexa Fluor® 488 Anti-Niemann Pick C1 antibody [EPR5209] (ab317983)
- Alexa Fluor® 594 Anti-Niemann Pick C1 antibody [EPR5209] (ab318025)
- Alexa Fluor® 555 Anti-Niemann Pick C1 antibody [EPR5209] (ab318067)
- Alexa Fluor® 647 Anti-Niemann Pick C1 antibody [EPR5209] (ab318108)
- Alexa Fluor® 750 Anti-Niemann Pick C1 antibody [EPR5209] (ab321075)
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab134113于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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Flow Cyt (Intra) |
1/200.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB | (1) |
1/2000 - 1/10000. Detects a band of approximately 180 kDa (predicted molecular weight: 142 kDa).
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IHC-P |
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ICC/IF | (1) |
1/70.
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说明 |
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Flow Cyt (Intra)
1/200. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/2000 - 1/10000. Detects a band of approximately 180 kDa (predicted molecular weight: 142 kDa). |
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC/IF
1/70. |
靶标
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功能
Involved in the intracellular trafficking of cholesterol. May play a role in vesicular trafficking in glia, a process that may be crucial for maintaining the structural and functional integrity of nerve terminals. -
疾病相关
Defects in NPC1 are the cause of Niemann-Pick disease type C1 (NPDC1) [MIM:257220]. A lysosomal storage disorder that affects the viscera and the central nervous system. It is due to defective intracellular processing and transport of low-density lipoprotein derived cholesterol. It causes accumulation of cholesterol in lysosomes, with delayed induction of cholesterol homeostatic reactions. Niemann-Pick disease type C1 has a highly variable clinical phenotype. Clinical features include variable hepatosplenomegaly and severe progressive neurological dysfunction such as ataxia, dystonia and dementia. The age of onset can vary from infancy to late adulthood. An allelic variant of Niemann-Pick disease type C1 is found in people with Nova Scotia ancestry. Patients with the Nova Scotian clinical variant are less severely affected. -
序列相似性
Belongs to the patched family.
Contains 1 SSD (sterol-sensing) domain. -
结构域
A cysteine-rich N-terminal domain and a C-terminal domain containing a di-leucine motif necessary for lysosomal targeting are critical for mobilization of cholesterol from lysosomes. -
翻译后修饰
Glycosylated. -
细胞定位
Late endosome membrane. Lysosome membrane. - Information by UniProt
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数据库链接
- Entrez Gene: 4864 Human
- Entrez Gene: 18145 Mouse
- Entrez Gene: 266732 Rat
- Omim: 607623 Human
- SwissProt: O15118 Human
- SwissProt: O35604 Mouse
- Unigene: 464779 Human
- Unigene: 715623 Human
see all -
别名
- Niemann Pick C1 protein precursor antibody
- Niemann Pick disease, type C1 antibody
- Niemann-Pick C1 protein antibody
see all
图片
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All lanes : Anti-Niemann Pick C1 antibody [EPR5209] (ab134113) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : NPC1 (Niemann Pick C1) knockout HAP1 whole cell lysate
Lane 3 : HEK293 whole cell lysate
Lane 4 : HepG2 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 142 kDaLanes 1 - 4: Merged signal (red and green). Green - ab134113 observed at 180 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab134113 was shown to specifically react with Niemann Pick C1 in wild-type HAP1 cells as signal was lost in NPC1 (Niemann Pick C1) knockout cells. Wild-type and NPC1 (Niemann Pick C1) knockout samples were subjected to SDS-PAGE. Ab134113 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (mouse neuroblastoma cell line) cells labeling Niemann Pick C1 with ab134113 at 1/70 followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500(green). Confocal image showing cytoplasmic staining on Neuro-2a cell line. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 and ab150120(AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 (red).
The negative controls are as follows:
-ve control 1 – ab134113 at 1/70 followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500.
-ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/1000 followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
Immunohistochemical staining of paraffin embedded human liver with purified ab134113 at a working dilution of 1 in 50. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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ab134113 staining Niemann Pick C1 in Neuro-2a (mouse neuroblastoma cell line) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde, permabilised with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/200. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.
Isoytype control: Rabbit monoclonal IgG (Black)
Unlabeled control: Cell without incubation with primary antibody and secondary antibody (Blue)
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Immunocytochemistry/ Immunofluorescence - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)Schultz et al Nat Commun. 2018; 9: 3671. Published online 2018 Sep 10. doi: 10.1038/s41467-018-06115-2
I1061T NPC1 traffics to autophagosomes
I1061T NPC1 fibroblasts were treated with vehicle or 100 nM bafilomycin A1 (Baf) for 24 h. Fixed cells were stained for LC3 (red), NPC1 (green), and DAPI (blue) then imaged by confocal microscopy. Co-localization is indicated by yellow color in the merged image. Scale bar = 25 μm.
Niemann Pick C1 (NPC1) was detected using ab134113 at 1/200 dilution.
From Figure 4b of Schultz et al.
Shultz et al Nat Commun. 2018; 9: 3671.Published online 2018 Sep 10. doi: 10.1038/s41467-018-06115-2
Reproduced under the Creative Commons Licence http://creativecommons.org/licenses/by/4.0/.
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Immunocytochemistry/ Immunofluorescence - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)Image from Cermak S et al., PLoS One. 2016;11(11):e0167428. Fig 3.; doi: 10.1371/journal.pone.0167428. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
Cathepsin B/L inhibition causes NPC disease-like cholesterol accumulation in SH-SY5Y cells.
Confocal microscopy of SH-SY5Y control and PADK treated cells. Cholesterol (filipin staining, white) and NPC1 (ab134113; green).
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All lanes : Anti-Niemann Pick C1 antibody [EPR5209] (ab134113) at 1/10000 dilution (purified)
Lane 1 : HepG2 (human liver hepatocellular carcinoma cell line) cell lysate
Lane 2 : THP-1 (human monocytic leukemia cell line) cell lysate
Lane 3 : HEK-293 (human epithelial cell line from embryonic kidney) cell lysate
Lane 4 : PC-3 (human prostate adenocarcinoma cell line) cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 142 kDa
Additional bands at: 180 kDa (possible glycosylated form)Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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All lanes : Anti-Niemann Pick C1 antibody [EPR5209] (ab134113) at 1/10000 dilution (purified)
Lane 1 : 3T3-L1 cell lysate
Lane 2 : L6 (rat skeletal muscle cell line) cell lysate
Lane 3 : Rat tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 142 kDa
Additional bands at: 180 kDa (possible glycosylated form)Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
Immunohistochemical staining of paraffin embedded mouse liver with purified ab134113 at a working dilution of 1 in 50. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
Immunohistochemical staining of paraffin embedded rat cerebellum with purified ab134113 at a working dilution of 1 in 50. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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All lanes : Anti-Niemann Pick C1 antibody [EPR5209] (ab134113) at 1/1000 dilution (unpurified)
Lane 1 : 3T3 L1 cell lysate
Lane 2 : L6 (rat skeletal muscle cell line) cell lysate
Lane 3 : HepG2 (human liver hepatocellular carcinoma cell line) cell lysate
Lane 4 : THP-1 (human monocytic leukemia cell line) cell lysate
Lane 5 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cell lysate
Lane 6 : PC-3 (human prostate adenocarcinoma cell line) cell lysate
Lane 7 : Rat liver lysate
Lane 8 : Rat brain lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution
Predicted band size: 142 kDa
Observed band size: 180 kDa why is the actual band size different from the predicted? -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Niemann Pick C1 antibody [EPR5209] (ab134113)
Immunohistochemical analysis of paraffin embedded human kidney tissue labelling Niemann Pick C1 with unpurified ab134113 at 1/50.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (51)
ab134113 被引用在 51 文献中.
- Nalbach K et al. Spatial proteomics reveals secretory pathway disturbances caused by neuropathy-associated TECPR2. Nat Commun 14:870 (2023). PubMed: 36797266
- Quelle-Regaldie A et al. Severe neurometabolic phenotype in npc1-/- zebrafish with a C-terminal mutation. Front Mol Neurosci 16:1078634 (2023). PubMed: 37008782
- Stewart CM et al. Sphingosine Kinases Promote Ebola Virus Infection and Can Be Targeted to Inhibit Filoviruses, Coronaviruses, and Arenaviruses Using Late Endocytic Trafficking to Enter Cells. ACS Infect Dis 9:1064-1077 (2023). PubMed: 37053583
- Kunkel TJ et al. The cholesterol transporter NPC1 is essential for epigenetic regulation and maturation of oligodendrocyte lineage cells. Nat Commun 14:3964 (2023). PubMed: 37407594
- Chiang YT et al. Cyclodextrin-Mediated Cholesterol Depletion Induces Adiponectin Secretion in 3T3-L1 Adipocytes. Int J Mol Sci 24:N/A (2023). PubMed: 37834165