重组Anti-INPPL1/SHIP-2抗体[EPR10955] (ab166916)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR10955] to INPPL1/SHIP-2
- Suitable for: 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-INPPL1/SHIP-2抗体[EPR10955]
参阅全部 INPPL1/SHIP-2 一抗 -
描述
兔单克隆抗体[EPR10955] to INPPL1/SHIP-2 -
宿主
Rabbit -
经测试应用
适用于: WB, IHC-P, ICC/IFmore details
不适用于: Flow Cyt or IP -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide within Human INPPL1/SHIP-2 aa 950-1050. The exact sequence is proprietary.
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阳性对照
- K562, HeLa whole cell lysate (ab150035); Human heart tissue; Human kidney tissue; HeLa 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. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading...
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纯度
Tissue culture supernatant -
克隆
单克隆 -
克隆编号
EPR10955 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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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ab166916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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WB |
1/1000 - 1/10000. Predicted molecular weight: 139 kDa.
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IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ICC/IF |
1/100 - 1/250.
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说明 |
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WB
1/1000 - 1/10000. Predicted molecular weight: 139 kDa. |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC/IF
1/100 - 1/250. |
靶标
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功能
Phosphatidylinositol (PtdIns) phosphatase that specifically hydrolyzes the 5-phosphate of phosphatidylinositol-3,4,5-trisphosphate (PtdIns(3,4,5)P3) to produce PtdIns(3,4)P2, thereby negatively regulating the PI3K (phosphoinositide 3-kinase) pathways. Plays a central role in regulation of PI3K-dependent insulin signaling, although the precise molecular mechanisms and signaling pathways remain unclear. While overexpression reduces both insulin-stimulated MAP kinase and Akt activation, its absence does not affect insulin signaling or GLUT4 trafficking. Confers resistance to dietary obesity. May act by regulating AKT2, but not AKT1, phosphorylation at the plasma membrane. Part of a signaling pathway that regulates actin cytoskeleton remodeling. Required for the maintenance and dynamic remodeling of actin structures as well as in endocytosis, having a major impact on ligand-induced EGFR internalization and degradation. Participates in regulation of cortical and submembraneous actin by hydrolyzing PtdIns(3,4,5)P3 thereby regulating membrane ruffling. Regulates cell adhesion and cell spreading. Required for HGF-mediated lamellipodium formation, cell scattering and spreading. Acts as a negative regulator of EPHA2 receptor endocytosis by inhibiting via PI3K-dependent Rac1 activation. Acts as a regulator of neuritogenesis by regulating PtdIns(3,4,5)P3 level and is required to form an initial protrusive pattern, and later, maintain proper neurite outgrowth. Acts as a negative regulator of the FC-gamma-RIIA receptor (FCGR2A). Mediates signaling from the FC-gamma-RIIB receptor (FCGR2B), playing a central role in terminating signal transduction from activating immune/hematopoietic cell receptor systems. Involved in EGF signaling pathway. Upon stimulation by EGF, it is recruited by EGFR and dephosphorylates PtdIns(3,4,5)P3. Plays a negative role in regulating the PI3K-PKB pathway, possibly by inhibiting PKB activity. Down-regulates Fc-gamma-R-mediated phagocytosis in macrophages independently of INPP5D/SHIP1. In macrophages, down-regulates NF-kappa-B-dependent gene transcription by regulating macrophage colony-stimulating factor (M-CSF)-induced signaling. May also hydrolyze PtdIns(1,3,4,5)P4, and could thus affect the levels of the higher inositol polyphosphates like InsP6. -
组织特异性
Widely expressed, most prominently in skeletal muscle, heart and brain. Present in platelets. Expressed in transformed myeloid cells and in primary macrophages, but not in peripheral blood monocytes. -
疾病相关
Defects in INPPL1 may be a cause of susceptibility to type 2 diabetes mellitus non-insulin dependent (NIDDM) [MIM:125853].
Note=Genetic variations in INPPL1 may be a cause of susceptibility to metabolic syndrome. Metabolic syndrome is characterized by diabetes, insulin resistance, hypertension, and hypertriglyceridemia is absent. -
序列相似性
Belongs to the inositol 1,4,5-trisphosphate 5-phosphatase family.
Contains 1 SAM (sterile alpha motif) domain.
Contains 1 SH2 domain. -
结构域
The SH2 domain interacts with tyrosine phosphorylated forms of proteins such as SHC1 or FCGR2A. It also mediates the interaction with p130Cas/BCAR1.
The NPXY sequence motif found in many tyrosine-phosphorylated proteins is required for the specific binding of the PID domain. -
翻译后修饰
Tyrosine phosphorylated by the members of the SRC family after exposure to a diverse array of extracellular stimuli such as insulin, growth factors such as EGF or PDGF, chemokines, integrin ligands and hypertonic and oxidative stress. May be phosphorylated upon IgG receptor FCGR2B-binding. Phosphorylated at Tyr-986 following cell attachment and spreading. Phosphorylated at Tyr-1162 following EGF signaling pathway stimulation. Phosphorylated at Thr-958 in response to PDGF. -
细胞定位
Cytoplasm > cytosol. Cytoplasm > cytoskeleton > actin patch. Membrane. Translocates to membrane ruffles when activated, translocation is probably due to different mechanisms depending on the stimulus and cell type. Partly translocated via its SH2 domain which mediates interaction with tyrosine phosphorylated receptors such as the FC-gamma-RIIB receptor (FCGR2B). Tyrosine phosphorylation may also participate in membrane localization. Insulin specifically stimulates its redistribution from the cytosol to the plasma membrane. Recruited to the membrane following M-CSF stimulation. - Information by UniProt
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数据库链接
- Entrez Gene: 3636 Human
- Entrez Gene: 16332 Mouse
- Entrez Gene: 65038 Rat
- Omim: 600829 Human
- SwissProt: O15357 Human
- SwissProt: Q6P549 Mouse
- SwissProt: Q9WVR3 Rat
- Unigene: 523875 Human
see all -
别名
- 4 antibody
- 5-trisphosphate 5-phosphatase 2 antibody
- 51C protein antibody
see all
图片
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All lanes : Anti-INPPL1/SHIP-2 antibody [EPR10955] (ab166916) at 1/1000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : INPPL1 knockout A549 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 139 kDa
Observed band size: 120-150 kDa why is the actual band size different from the predicted?Western blot: Anti-INPPL1 antibody [EPR10955] (ab166916) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab166916 was shown to bind specifically to INPPL1. A band was observed at 120-150 kDa in wild-type A549 cell lysates with no signal observed at this size in INPPL1 knockout cell line. To generate this image, wild-type and INPPL1 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: INPPL1/SHIP-2 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: K562 whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab166916 observed at 139 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab166916 was shown to specifically react with INPPL1/SHIP-2 in wild-type HAP1 cells. No band was observed when INPPL1/SHIP-2 knockout samples were examined. Wild-type and INPPL1/SHIP-2 knockout samples were subjected to SDS-PAGE. Ab166916 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10,000 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 1/10,000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-INPPL1/SHIP-2 antibody [EPR10955] (ab166916)
Immunohistochemical analysis of paraffin-embedded Human heart tissue labeling INPPL1/SHIP-2 with ab166916 at 1/100 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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All lanes : Anti-INPPL1/SHIP-2 antibody [EPR10955] (ab166916) at 1/1000 dilution
Lane 1 : K562 cell lysate
Lane 2 : HeLa cell lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 139 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-INPPL1/SHIP-2 antibody [EPR10955] (ab166916)
Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling INPPL1/SHIP-2 with ab166916 at 1/100 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunofluorescent analysis of HeLa cells labeling INPPL1/SHIP-2 with ab166916 at 1/100 dilution.
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (1)
ab166916 被引用在 1 文献中.
- Iguchi A et al. INPP5D modulates TREM2 loss-of-function phenotypes in a β-amyloidosis mouse model. iScience 26:106375 (2023). PubMed: 37035000