使用敲除细胞株进行验证

Anti-Insulin degrading enzyme / IDE抗体(ab32216)

概述

  • 产品名称Anti-Insulin degrading enzyme / IDE抗体
    参阅全部 Insulin degrading enzyme / IDE 一抗
  • 描述
    兔多克隆抗体to Insulin degrading enzyme / IDE
  • 经测试应用适用于: WB, IHC-P, IHC-FoFr, ICC/IF, IHC-Frmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
    预测可用于: Cow, Cat, Dog
  • 免疫原

    Synthetic peptide derived from within residues 950 to the C-terminus of Human Insulin degrading enzyme/ IDE.

    (Peptide available as ab32215.)

  • 阳性对照
    • Mouse and Rat brain lysate. Mouse and Rat hippocampus lysate

性能

应用

Our Abpromise guarantee covers the use of ab32216 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 118 kDa (predicted molecular weight: 118 kDa).Can be blocked with Human Insulin degrading enzyme / IDE peptide (ab32215).
IHC-P 1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-FoFr 1/100.
ICC/IF 1/500.
IHC-Fr 1/500.

靶标

  • 功能Plays a role in the cellular breakdown of insulin, IAPP, glucagon, bradykinin, kallidin and other peptides, and thereby plays a role in intercellular peptide signaling. Degrades amyloid formed by APP and IAPP. May play a role in the degradation and clearance of naturally secreted amyloid beta-protein by neurons and microglia.
  • 序列相似性Belongs to the peptidase M16 family.
  • 翻译后修饰The N-terminus is blocked.
  • 细胞定位Cytoplasm. Cell surface. Present at the cell surface of neuron cells. The membrane-associated isoform is approximately 5 kDa larger than the known cytosolic isoform.
  • Information by UniProt
  • 数据库链接
  • 别名
    • Abeta-degrading protease antibody
    • FLJ35968 antibody
    • Ide antibody
    • IDE_HUMAN antibody
    • Insulin protease antibody
    • Insulin-degrading enzyme antibody
    • Insulinase antibody
    • Insulysin antibody
    • OTTHUMP00000020097 antibody
    see all

Anti-Insulin degrading enzyme / IDE antibody 图像



  • Predicted band size : 118 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: Insulin degrading enzyme / IDE knockout HAP1 cell lysate (20 µg)
    Lane 3: K562 cell lysate (20 µg)
    Lane 4: HepG2 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab32216 observed at 120 kDa. Red - loading control, ab8245, observed at 37 kDa.
    ab32216 was shown to specifically react with Insulin degrading enzyme / IDE when Insulin degrading enzyme / IDE knockout samples were used. Wild-type and Insulin degrading enzyme / IDE knockout samples were subjected to SDS-PAGE. ab32216 and ab8245 (loading control to GAPDH) were diluted at 1 μg/ml and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) ab216776 secondary antibodies at 1/10,000 dilution for 1 h at room temperature before imaging.

  • Immunofluorescent staining for Insulin degrading enzyme/IDE in rat brain rat hippocampus using Rabbit polyclonal to Insulin degrading enzyme/IDE (ab32216). . The staining is located in the neuronal soma and is finely punctuated. The picture was acquired using the X20 objective. Protocol details: Rats were intracardially perfused with 4% paraformaldehyde. Whole brain tissue was post-fixed overnight in the same fixative, and cryoprotected in 20% sucrose and frozen in OCT. 30µm coronal sections were cut by cryostat for use in fre floating IHC. Primary antibody ab32216 was incubated overnight at 1/100 at room temperature. Secondary antibody Alexa fluor 488 1/1000 was incubated for 2 hours at room temperature.
  • Image courtesy of Human Protein Atlas

    ab32216 staining human Gall bladder. The paraffin embedded human skin tissue was incubated with ab32216 (1/50 dilution) for 30 mins at room temperature. The staining is predominantly cytoplasmic although some nuclear staining can be seen. Antigen retrieval was performed by heat induction in citrate buffer pH 6. Ab32216 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines.

    Further results for

  • ab32216 staining cultured rat OCP by ICC/IF. The cultured cells were fixed with paraformaldehyde and blocked with 10% donkey serum for 30 minutes at 24°C. The cultured cellswere then stained with ab322161 at 1/500 in 0.3% TritonX with 0.1% PBS and 10% donkey serum for 4h at 24°C. An Alexa Fluro 488 donkey anti-rabbit polyclonal antibody at 1/1000 was used as the secondary antibody. Nuclei were stained with 1.43µM Hoechst and can be observed in blue.

    See Abreview

  • All lanes : Anti-Insulin degrading enzyme / IDE antibody (ab32216) at 1 µg/ml

    Lane 1 : Mouse Brain at 20 µg/ml
    Lane 2 : Brain (Rat) Whole Cell Lysate - normal tissue at 20 µg
    Lane 3 : Mouse Hippocampus Lysate at 20 µg
    Lane 4 : Rat Hippocampus Lysate at 20 µg

    Secondary
    Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 118 kDa
    Observed band size : 118 kDa

Anti-Insulin degrading enzyme / IDE antibody (ab32216)参考文献

This product has been referenced in:
  • Ries M  et al. The anti-inflammatory Annexin A1 induces the clearance and degradation of the amyloid-ß peptide. J Neuroinflammation 13:234 (2016). Read more (PubMed: 27590054) »
  • Santos RS  et al. Lacking of estradiol reduces insulin exocytosis from pancreatic ß-cells and increases hepatic insulin degradation. Steroids 114:16-24 (2016). WB . Read more (PubMed: 27192429) »

See all 10 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Brainstem and kidney)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Specification Brainstem and kidney
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

提交于 Sep 26 2016

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Rat Tissue sections (brain)
Specification brain
Fixative Paraformaldehyde
Antigen retrieval step Other
Permeabilization No
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 8% · Temperature: 37°C
Username

Abcam user community

Verified customer

提交于 Feb 28 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Rat Cell (OPC culture)
Specification OPC culture
Fixative Paraformaldehyde
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C
Username

Dr. Ruma Raha-Chowdhury

Verified customer

提交于 Feb 10 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Rat Tissue sections (Muscle)
Specification Muscle
Fixative Paraformaldehyde
Permeabilization Yes - 0.3% TritonX in 0.1% PBS
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Username

Dr. Ruma Raha-Chowdhury

Verified customer

提交于 Jun 02 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Rat Tissue lysate - whole (Brain, liver tissues,)
Loading amount 20 µg
Specification Brain, liver tissues,
Gel Running Conditions Reduced Denaturing (12 % Tris Tricine gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Dr. Ruma Raha-Chowdhury

Verified customer

提交于 May 20 2011

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