Anti-Insulin Receptor beta抗体[C18C4] (ab69508)

概述

  • 产品名称
    Anti-Insulin Receptor beta抗体[C18C4]
    参阅全部 Insulin Receptor beta 一抗
  • 描述
    小鼠单克隆抗体[C18C4] to Insulin Receptor beta
  • 宿主
    Mouse
  • 经测试应用
    适用于: Flow Cyt, IHC-P, WBmore details
  • 种属反应性
    与反应: Mouse, Rat, Sheep, Rabbit, Hamster, Cow, Human, Pig
  • 免疫原

    Recombinant cytoplasmic domain of human Insulin Receptor beta

  • 阳性对照
    • MCF-7, Rat-2 and L-929 cells

性能

应用

Our Abpromise guarantee covers the use of ab69508 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 95 kDa (predicted molecular weight: 95 kDa). (developed by ECL)

靶标

  • 相关性
    Insulin receptor mediates the biological activities of insulin by regulating multiple signaling pathways through activation of a series of phosphorylation cascades. The human insulin receptor is a heterotetrameric membrane glycoprotein consisting of disulfide-linked subunits in a ß-a-a-ß configuration. The ß-subunit (95kDa) possesses a single transmembrane domain with tyrosine kinase acivity, whereas the a-subunit (135kDa) is completely extracellular. The alpha subunits each contain insulin binding sites and are entirely extracellular in localization. The beta subunits each possess an extracellular domain, a single transmembrane domain, and a cytoplasmic tyrosine kinase domain. Binding of insulin to the alpha subunits induces a conformation change in the receptor which activates the kinase domain, stimulating tyrosine autophosphorylation of the receptor and tyrosine phosphorylation of at least five different insulin receptor substrates designated IRS-1-4, and Shc.
  • 细胞定位
    Membrane; Single pass type I membrane protein.
  • 数据库链接
  • 别名
    • CD220 antibody
    • HHF5 antibody
    • HIR B antibody
    • INSR antibody
    • Insulin receptor antibody
    • Insulin receptor subunit beta antibody
    • IR antibody
    see all

图片

  • All lanes : Anti-Insulin Receptor beta antibody [C18C4] (ab69508) at 1/1000 dilution

    Lane 2 : HeLa lysate (Human epithelial cell line from cervix adenocarcinoma)
    Lane 3 : MCF-7 lysate (Human breast adenocarcinoma cell line)
    Lane 4 : PC-12 lysate (Rat adrenal gland pheochromocytoma cell line)
    Lane 5 : 3T3-L1 lysate (Mouse embryonic fibroblast cell line)

    Developed using the ECL technique.

    Predicted band size: 95 kDa
    Observed band size: 98 kDa (why is the actual band size different from the predicted?)

  • IHC image of ab69508 staining in human normal kidney formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab69508, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Overlay histogram showing HepG2 cells stained with ab69508 (red line). The cells were fixed with 80% methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab69508, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.

文献

This product has been referenced in:

See all 9 Publications for this product

客户评价及客户问答

Abcam has not validated the combination of species/application used in this Abreview.
Application
Flow Cytometry
Sample
Mouse Cell (HepG2 Human liver cells)
Permeabilization
No
Gating Strategy
Population was gated with the FSC-SSC diagram
Specification
HepG2 Human liver cells
Preparation
Cell harvesting/tissue preparation method: Steps: Detach cells with enzymatic treatment (Accutase) and proceed to cell counting and viability estimation Wash the cells with ice-cold PBS 1X Repeat the step For native cells : resuspend the cells in ice-cold PBS-BSA 1% to obtain a 2 millions of/ml suspension, distribute 100µl / well (200.000 cells/96 well) and incubate/block on ice during 15min For methanol-fixed cells: resuspend the cells in ice-cold PBS to obtain a 2 millions of/ml suspension, distribute 100µl / well (200.000 cells/96 well), centrifuge the plate at 750g, 4°C, 1min to discard the supernatant, resuspend each well with 100µl of ice-cold methanol 80% in PBS, incubate 10min on ice, make 3 washes of cells with 200µl of ice-cold PBS, resuspend in 100µl of ice-cold PBS-BSA 1%, and incubate/block on ice during 15min For PFA-fixed cells: resuspend the cells in ice-cold PBS to obtain a 2 millions of/ml suspension, distribute 100µl / well (200.000 cells/96 well), centrifuge the plate at 750g, 4°C, 1min to discard the supernatant, resuspend each well with 100µl of ice-cold PFA 2% in PBS, incubate 10min on ice, make 3 washes of cells with 200µl of ice-cold PBS, resuspend in 100µl of ice-cold PBS-BSA 1%, and incubate/block on ice during 15min Centrifuge the plates at 750g, 4°C, 1min to discard the supernatant Protocole for primary antibody with secondary detection Add primary antibody at 0.025µM in 100µl of PBS-BSA 1% Incubate on ice during 30min Wash the cells with 200µl of ice-cold PBS 1X Repeat the step (x2) Add secondary antibody at 0.025µM in 100µl of PBS-BSA 1% Incubate on ice during 30min Wash the cells with 200µl of ice-cold PBS 1X Repeat the step (x2) Resuspend the cells with 200µl of ice-cold PBS 1X Analyze the cells by flow cytometry (10.000 events in a viability gate)
Sample buffer: Classical PBS-BSA 1 %
Fixation
Comparison of antibody binding on PFA fixed cells, methanol fixed cells and non fixed native cells
Username

Abcam user community

Verified customer

提交于 Mar 15 2018

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Bos taurus Tissue lysate - whole (Liver)
Gel Running Conditions
Reduced Denaturing (4-15% gradient)
Loading amount
20 µg
Specification
Liver
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Julie Kim

Verified customer

提交于 Apr 13 2017



According to the Uniprot homepage the alpha chain is located from amino acid 28-758, whereas the beta chain is from aa 763-1382:

http://www.uniprot.org/uniprot/P06213

ab69508 has its immunogen in the cytoplasmic domain of ...

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I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

I am at...

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Thank you for your enquiry. I am sorry to hear this vial of ab52167 is giving you trouble. We don't have a specificIHC-P protocol available for this antibody. But If you could, please complete the formbelow so I can get an idea of your protocol and dat...

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Merci de votre intérêt pour les anticorps Abcam. Oui, je recommanderais ab37796 pour le marquage de la glucokinase et ab69508 pour le récepteur insuline beta. Par contre, pour le marquage de PCK1 je recommanderais plutôt un anticorps test...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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