Anti-Insulin Receptor (phospho Y972)抗体
参阅全部 Insulin Receptor 一抗
兔多克隆抗体to Insulin Receptor (phospho Y972)
Rabbit
In some cell systems ab5678 has been shown to cross-react with IGF1R pY950 (75% homologous).
适用于: ICC/IF, WBmore details
与反应: Human
预测可用于: Mouse, Rat
Synthetic peptide corresponding to Human Insulin Receptor (phospho Y972).
IF: Insulin treated MCF7 cells. WB: CHO-T (Chinese hamster ovary cell line) cells transfected with a vector encoding the human insulin receptor and stimulated with insulin.
Biological actions of insulin are mediated by the Insulin Receptor (IR), a receptor tyrosine kinase that regulates multiple signaling pathways through activation of a series of phosphorylation cascades. The IR is a heterotetrameric protein consisting of two ligand-binding alpha subunits and two beta subunits that each contain a tyrosine kinase domain. Insulin binding to the extracellular domain leads to autophosphorylation of the receptor and activation of the intrinsic tyrosine kinase activity, which allows appropriate substrates to be phosphorylated. Tyrosine 972 is in the juxtamembrane Asn-Pro- Glu-Tyr (NPEY) motif. Phosphorylation of IR tyrosine 972 is required for the binding and/or phosphorylation of the adapter protein Shc, the PTB domain, IRS-1, PI3 kinase, and the Suppressor of Cytokine Signaling (SOCS).
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Liquid
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
pH: 7.30
Preservative: 0.05% Sodium azide
Constituents: PBS, 1% BSA, 50% Glycerol
浓度
50 µl 浓度为 0.8 mg/ml
Immunogen affinity purified
The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated Insulin Receptor (IR). The final product is generated by affinity chromatography using an IR-derived peptide phosphorylated at tyrosine 972.
Biological actions of insulin are mediated by the Insulin Receptor (IR), a receptor tyrosine kinase that regulates multiple signaling pathways through activation of a series of phosphorylation cascades. The IR is a heterotetrameric protein consisting of two ligand-binding alpha subunits and two beta subunits that each contain a tyrosine kinase domain. Insulin binding to the extracellular domain leads to autophosphorylation of the receptor and activation of the intrinsic tyrosine kinase activity, which allows appropriate substrates to be phosphorylated. Tyrosine 972 is in the juxtamembrane Asn-Pro- Glu-Tyr (NPEY) motif. Phosphorylation of IR tyrosine 972 is required for the binding and/or phosphorylation of the adapter protein Shc, the PTB domain, IRS-1, PI3 kinase, and the Suppressor of Cytokine Signaling (SOCS).
多克隆
IgG
Abpromise™承诺保证使用ab5678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 |
|---|---|---|
| ICC/IF | 1/100 - 1/500. | |
| WB | (2) | 1/1000. Detects a band of approximately 110 kDa. |
Entrez Gene: 3643 Human
Entrez Gene: 16337 Mouse
Omim: 147670 Human
SwissProt: P06213 Human
SwissProt: P15208 Mouse
Unigene: 465744 Human
Unigene: 268003 Mouse
Unigene: 9876 Rat
CD220 antibody
HHF5 antibody
human insulin receptor antibody
Insr antibody
INSR_HUMAN antibody
Insulin receptor subunit beta antibody
IR 1 antibody
IR antibody
IR-1 antibody
IR1 antibody
Immunocytochemistry/ Immunofluorescence - Anti-Insulin Receptor (phospho Y972) antibody (ab5678)
Immunofluorescence analysis of insulin treated MCF7 cells labelling Insulin Receptor (phospho Y972) (Panel a: green) using ab5678 at 2µg/mL in 1% BSA for 3 hours at room temperature, followed by Alexa Fluor 488® Goat Anti-Rabbit IgG Secondary Antibody at 1/400 dilution for 30 minutes at room temperature. Panel b:Nuclei were stained with DAPI (blue). Panel c: F-actin was stained with Alexa Fluor 594® Phalloidin (red). Panel d: Merged image showing membrane localization. Panel e: Untreated MCF7 cells. Panel f: Control, no primary antibodyl. The images were captured at 20X magnification.
Prior antibody incubation, MCF7 (human breast adenocarcinoma cell line) cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature, followed by treatment with 100nM of insulin for 5 min. Assay was done on 70% confluent log phase MCF7 cells.
Western blot - Anti-Insulin Receptor (phospho Y972) antibody (ab5678)
All lanes : Anti-Insulin Receptor (phospho Y972) antibody (ab5678) at 1/1000 dilution (2 hours at room temperature in a 3% BSA-TBST buffer)
Lane 1 : Unstimulated (-), CHO-T transfected with insulin receptor containing vector whole cell extract with 5% BSA-TBST buffer for one hour at room temperature
Lanes 2-5 : Stimulated (+) with 50 nM insulin for 5 minutes, CHO-T transfected with insulin receptor containing vector whole cell extract with 5% BSA-TBST buffer for one hour at room temperature
Secondary
All lanes : Goat F (ab')2 anti-rabbit IgG HRP conjugate
Upregulation and Antibody-Peptide Competition:
Prior primary antibody incubation:
1 and 2 - no peptide;
3 - non-phosphorylated peptide corresponding to the phosphopeptide immunogen;
4 - generic phosphotyrosine-containing peptide;
5 - phosphopeptide immunogen.
SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF.
The data show that only the phosphopeptide corresponding to ab5678 completely blocks the antibody signal, demonstrating the specificity of the antibody.
The data also show up-regulation of the signal upon stimulation with insulin in this cell system.
