Anti-Niemann Pick C2 抗体 [EPR19993-145-1] - BSA and Azide free (ab251560)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19993-145-1] to Niemann Pick C2 - BSA and Azide free
- Suitable for: ICC/IF, IHC-P, Flow Cyt (Intra), WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-Niemann Pick C2 antibody [EPR19993-145-1] - BSA and Azide free
Niemann Pick C2 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR19993-145-1] to Niemann Pick C2 - BSA and Azide free -
由来種
Rabbit -
アプリケーション
適用あり: ICC/IF, IHC-P, Flow Cyt (Intra), WBmore details -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
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ポジティブ・コントロール
- WB: Human fetal kidney and spleen lysates; Mouse kidney lysate; Rat kidney and spleen lysates; HEK-293T, HepG2, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates. IHC-P: Human epididymis and testis tissues, human breast carcinoma; Mouse and rat kidney tissues. Flow Cyt (intra): HepG2 and NIH/3T3 cells.
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特記事項
ab251560 is the carrier-free version of ab218192.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
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.
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.
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.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
バッファー
pH: 7.2
Constituent: PBS -
キャリア・フリー
はい -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EPR19993-145-1 -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Products
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab251560の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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ICC/IF |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 16-18 kDa (predicted molecular weight: 17 kDa).
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特記事項 |
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ICC/IF
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 16-18 kDa (predicted molecular weight: 17 kDa). |
ターゲット情報
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機能
May be involved in the regulation of the lipid composition of sperm membranes during the maturation in the epididymis. -
組織特異性
Epididymis. -
関連疾患
Defects in NPC2 are the cause of Niemann-Pick disease type C2 (NPDC2) [MIM:607625]. 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 C2 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. -
配列類似性
Belongs to the NPC2 family. -
細胞内局在
Secreted. - Information by UniProt
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参照データベース
- Entrez Gene: 10577 Human
- Entrez Gene: 67963 Mouse
- Entrez Gene: 286898 Rat
- Omim: 601015 Human
- SwissProt: P61916 Human
- SwissProt: Q9Z0J0 Mouse
- Unigene: 433222 Human
- Unigene: 282556 Mouse
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別名
- EDDM1 antibody
- Epididymal protein 1 antibody
- Epididymal secretory protein antibody
see all
画像
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All lanes : Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192) at 1/5000 dilution
Lane 1 : Human fetal kidney lysate
Lane 2 : Mouse kidney lysate
Lane 3 : Rat kidney lysate
Lane 4 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 5 : Rat spleen lysate
Lane 6 : Human spleen lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 17 kDa
Observed band size: 16-18 kDa why is the actual band size different from the predicted?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.
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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. -
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).
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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. -
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. -
All lanes : Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192) at 1/2000 dilution
Lane 1 : C6 (Rat glial tumor cell line) whole cell lysate
Lane 2 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 4 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 17 kDa
Observed band size: 16-18 kDa why is the actual band size different from the predicted?
Exposure time: 10 secondsThis 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.
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All lanes : Anti-Niemann Pick C2 antibody [EPR19993-145-1] (ab218192) 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
Lane 2 : NPC2 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Lane 3 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 17 kDa
Observed band size: 16-18 kDa why is the actual band size different from the predicted?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.
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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.
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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.
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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.
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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.
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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.
プロトコール
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
データシートおよび資料
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Datasheet download
Certificate of Compliance
参考文献 (0)
ab251560 は論文での使用が確認できていません。