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使用敲除细胞株进行验证

Anti-Insulin degrading enzyme / IDE抗体(ab32216)

  • Datasheet
  • SDS
Reviews (9)Q&A (5)References (82)

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Western blot - Anti-Insulin degrading enzyme / IDE antibody (ab32216)
  • Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Insulin degrading enzyme / IDE antibody (ab32216)
  • Western blot - Anti-Insulin degrading enzyme / IDE antibody (ab32216)

Key features and details

  • Rabbit polyclonal to Insulin degrading enzyme / IDE
  • Suitable for: WB, IHC-FoFr
  • Knockout validated
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG

选择批间可重复性更高的重组抗体

Product image
Anti-Insulin degrading enzyme / IDE antibody [EPR6098(2)] (ab133561)
  • 研究可靠 —— 各批次间结果一致且可重复
  • 长期批量供应 —— 采用重组技术,可实现快速生产
  • 首次实验即可成功 —— 经过大量验证确认了特异性
  • 符合伦理标准 —— 产品不含动物成分

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
  • 宿主

    Rabbit
  • 特异性

    Replenishment batches of our polyclonal antibody, ab32216 are tested in WB. Previous batches were additionally validated in IHC-FoFr. This application is still expected to work and is covered by our Abpromise guarantee. You may also be interested in our alternative recombinant antibody, ab133561.

  • 经测试应用

    适用于: WB, IHC-FoFrmore 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. This information is proprietary to Abcam and/or its suppliers.

  • 常规说明

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

性能

  • 形式

    Liquid
  • 存放说明

    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.
  • 存储溶液

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: 98.98% PBS, 1% BSA

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • 纯度

    Immunogen affinity purified
  • 克隆

    多克隆
  • 同种型

    IgG
  • 研究领域

    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Alzheimer's disease
    • Other
    • Signal Transduction
    • Growth Factors/Hormones
    • Insulin / Insulin-like
    • Neuroscience
    • Neurology process
    • Metabolism
    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteolytic enzymes
    • Metalloprotease
    • Insulysin
    • Cardiovascular
    • Atherosclerosis
    • Diabetes associated
    • Metabolism
    • Types of disease
    • Neurodegenerative disease
    • Metabolism
    • Types of disease
    • Diabetes
    • Metabolism
    • Types of disease
    • Cancer
    • Metabolism
    • Types of disease
    • Heart disease

相关产品

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human Insulin degrading enzyme / IDE protein (ab152467)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab32216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
WB (2)
Use a concentration of 1 µg/ml. Detects a band of approximately 118 kDa (predicted molecular weight: 118 kDa).
IHC-FoFr (2)
1/100.
说明
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 118 kDa (predicted molecular weight: 118 kDa).
IHC-FoFr
1/100.

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.
  • Target information above from: UniProt accession P14735 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 数据库链接

    • Entrez Gene: 523752 Cow
    • Entrez Gene: 477768 Dog
    • Entrez Gene: 3416 Human
    • Entrez Gene: 15925 Mouse
    • Entrez Gene: 25700 Rat
    • Omim: 146680 Human
    • SwissProt: Q24K02 Cow
    • SwissProt: P14735 Human
    • SwissProt: Q9JHR7 Mouse
    • SwissProt: P35559 Rat
    • Unigene: 500546 Human
    • Unigene: 28366 Mouse
    • Unigene: 45029 Rat
    see all
  • 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

图片

  • Western blot - Anti-Insulin degrading enzyme / IDE antibody (ab32216)
    Western blot - Anti-Insulin degrading enzyme / IDE antibody (ab32216)

    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 in wild-type HAP1 cells. No band was observed when Insulin degrading enzyme / IDE knockout samples were examined. Wild-type and Insulin degrading enzyme / IDE knockout samples were subjected to SDS-PAGE. ab32216 and ab8245 (loading control to GAPDH) were diluted at 1μg/ml and 1/10,000 respectively and incubated overnight at 4°C. 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 1/10,000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Insulin degrading enzyme / IDE antibody (ab32216)
    Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Insulin degrading enzyme / IDE antibody (ab32216)This image is courtesy of Sophie Pezet, CNRS, Paris, France
    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.
  • Western blot - Anti-Insulin degrading enzyme / IDE antibody (ab32216)
    Western blot - Anti-Insulin degrading enzyme / IDE antibody (ab32216)
    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
    All lanes : 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

实验方案

  • Immunohistochemistry protocols
  • Western blot protocols

Click here to view the general protocols

数据表及文件

  • SDS download

  • Datasheet download

    Download

文献 (82)

发表研究结果有使用 ab32216?请让我们知道,以便我们可以引用本数据表中的参考文章。

ab32216 被引用在 82 文献中.

  • Soares GM  et al. Sleeve Gastrectomy-Induced Weight Loss Increases Insulin Clearance in Obese Mice. Int J Mol Sci 24:N/A (2023). PubMed: 36675244
  • Espinosa-Jiménez T  et al. A novel rhein-huprine hybrid ameliorates disease-modifying properties in preclinical mice model of Alzheimer's disease exacerbated with high fat diet. Cell Biosci 13:52 (2023). PubMed: 36895036
  • Karahan H  et al. The effect of Abi3 locus deletion on the progression of Alzheimer's disease-related pathologies. Front Immunol 14:1102530 (2023). PubMed: 36895556
  • Zhai S  et al. The day-night differences in cognitive and anxiety-like behaviors of mice after chronic sleep restriction. FASEB J 37:e23034 (2023). PubMed: 37341989
  • Naia L  et al. Mitochondrial hypermetabolism precedes impaired autophagy and synaptic disorganization in App knock-in Alzheimer mouse models. Mol Psychiatry 28:3966-3981 (2023). PubMed: 37907591
View all Publications for this product

客户评价及客户问答

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1-10 of 14 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Insulin degrading enzyme / IDE antibody

Inconclusive
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MR. Carl Hobbs

Verified customer

提交于 Sep 26 2016

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Insulin degrading enzyme / IDE antibody

Excellent
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Feb 28 2012

Immunocytochemistry/ Immunofluorescence abreview for Anti-Insulin degrading enzyme / IDE antibody

Excellent
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Ruma Raha-Chowdhury

Verified customer

提交于 Feb 10 2012

Immunohistochemistry (Frozen sections) abreview for Anti-Insulin degrading enzyme / IDE antibody

Excellent
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Ruma Raha-Chowdhury

Verified customer

提交于 Jun 02 2011

Western blot abreview for Anti-Insulin degrading enzyme / IDE antibody

Excellent
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Ruma Raha-Chowdhury

Verified customer

提交于 May 20 2011

Question

I would like to order ab133561 as a replacement for ab32216. Please can you tell me how I do this - is there a code to enter when ordering?

Thanks

Read More

Abcam community

Verified customer

Asked on Nov 30 2012

Answer

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

I am sorry this antibody did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued an aternative free of charge replacement of ab133561 with the order number 1213991.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let me know.

I wish you the best of luck with your research.

Read More

Abcam Scientific Support

回复于 Nov 30 2012

Question

Thank you for your reply; I would be happy to try another Abcam antibody for the same protein. Could you tell me if you recommend ab133561 for human WB? Would this also be covered by the guarantee if (in the unlikely event) it fails to work?

Many thanks

Read More

Abcam community

Verified customer

Asked on Nov 29 2012

Answer

Thank you for your reply.

I would like to reassure you that we monitor feedback closely on a weekly basis and we are not currently concerned about the general quality of ab32216 IDE antibody or this batch. Regrettably, I can suggest you have received a bad vial on this occasion.

Therefore, we would be pleased to provide a replacement from the same antibody, which will still be covered by our guarantee. Alternatively, I fully understand your concerns and if you prefer an alternative replacement of ab133561 or a credit note in this case, I will be pleased to arrange this for you.

I can confirm that ab133561 IDE antibody is tested and guaranteed for both human samples and WB. All tested applications and species covered by the 6 month guarantee are listed on the individual datasheets. The replacement vial will also be covered by the guarantee.

I hope this will be helpful. I look forward to hearing from you with details of how you would like to proceed.

Read More

Abcam Scientific Support

回复于 Nov 29 2012

Question

Thank you for your reply, however I'm afraid I don't quite understand your answer and there may be some confusion here. The stable transfection was of choline transporter-1 and not IDE, and I am trying to determine the endogenous IDE levels. The same antibody was used successfully for ICC and for loading controls, I have attached an image of pan-PKB.

Best wishes

Read More

Abcam community

Verified customer

Asked on Nov 28 2012

Answer

Thank you for your message and for providing this further information.

I apologize for the confusion, this is clearer to me now. I am sorry to hear the suggestions made have not improved the results on this occasion. Unfortunately, reviewing the details I am uncertain as to why this antibody is not working in WB for you when it is giving successful results in ICC.

I appreciate the time you have spent on these experiments and would be pleased to arrange a free of charge replacement or credit note in compensation.

I look forward to hearing from you with details of how you would like to proceed.

Read More

Abcam Scientific Support

回复于 Nov 28 2012

Question

Order Details
Antibody code: ab32216

Problem
Choose: No signal

Lot number GR1623

Purchase order number #####
or preferably Abcam order number:

General Information
Antibody storage conditions (temperature/reconstitution etc) Aliquoted and stored -20

Description of the problem (high background, wrong band size, more bands, no band etc.) No bands

Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.) Human cell extract (SH-SY5Y) stable transformation of choline transporter 1.

Sample preparation (Buffer/Protease inhibitors/Heating sample etc.) Lysis buffer: 50mM Tris-HCl pH7.5, 150mM NaCl,1% Triton X-100, Protease inhibitors: 1mM AEBSF, 10ug/ml leupeptin, 25ug/ml aprotinin, 10ug/ml pepstatin A, 700 Units DNAse. Sample buffer (3x): 6% SDS, 0.1875M Tris-HCl pH6.8, 30% Glycerol, 0.015% Bromophenol blue, 7.5% 2-mercaptoethanol. Sample heated 10 mins 55 deg C.

Amount of protein loaded 50 micrograms

Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.) Reducing (2- mercaptoethanol included in sample buffer), 7.5%

Transfer and blocking conditions (Buffer/time period, Blocking agent etc.) semi-dry transfer (Bio Rad semi-dry transfer cell), PVDF membrane (pre-wet in methanol), Transfer buffer: 14.5g/L Tris-HCl, 7.25g/L Glycine, 20% methanol. Blocking: 8% milk in washing buffer, 1hr. Washing buffer: 1x phoshphate buffered saline pH 7.4, 0.15% Triton X-100.

Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) ab32216, rabbit polyclonal to Insulin Degrading Enzyme. Diluted 1/1000 in 8% milk in 1xPBS + 0.15% Triton X-100, overnight, 4 deg C. Washed 2x 30 seconds + 3x 10 minutes in washing buffer.

Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) goat anti-rabbit-HRP (IgG). 1/10,000 in 1x PBS +0.15% Triton X-100, 1hr room temperature. Washed 2x 30 seconds + 3x 10 minutes in washing buffer.

Detection method (ECL, ECLPlus etc.) ECL

Positive and negative controls used (please specify) Positive controls = successful detection of other antibodies e.g. phospho-PKB (Cell Signalling #4060). Additionally, detection of signal with immunohistochemistry in same cell line.

Optimization attempts (problem solving)
How many times have you tried the Western? Twice. First attempt was after blot had been stripped three times, so repeated with fresh (unstripped) blot.

Have you run a "No Primary" control?
No

Do you obtain the same results every time?
Yes
e.g. are the background bands always in the same place?

What steps have you altered?

Additional Notes:

Image:
We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to asess the results.

Read More

Abcam community

Verified customer

Asked on Nov 26 2012

Answer

Thank you for taking the time to complete our questionnaire.

The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

Reviewing this case, I would like to offer some suggestions to help optimize the results. I would also appreciate if you can confirm some further details:

1. There are inherent difficulties with antibody detection of recombinant tagged proteins that I can recommend considering. This may be a biological artifact rather than a problem with the antibody. Could you confirm that the protein transfected is full length and includes the immunogen sequence? Also, if the recombinant IDE is tagged, it is possible that the tag is preventing access to the antibody. Could you confirm at which region of the protein the tag has been placed?

2. Could you confirm the species from which the recombinant IDE gene was taken?

3. Could you confirm how the stability of transfection has been assessed?

4. I can recommend that due to the points described above, it is important to use an endogenous positive control sample. For example, this antibody has been used successfully on mouse and rat brain lysate.

5. Has the transfer of protein to the membrane and quality of the sample been assessed using a loading control?

6. Was the same antibody used for detection of a signal in the ICC experiments?

I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details. I hope we can resolve this case for you as soon as possible.

Read More

Abcam Scientific Support

回复于 Nov 26 2012

Question

Hello,

I recently purchased ab32216 Anti-Insulin degrading enzyme. Although it is listed to react with human, I cannot get it to work in Western blot for my human cell line (SY5Y). I was wondering if you could help me trouble-shoot this? I applied the primary and secondary (HRP conjugated enzyme) at the recommended concentrations to 50ug of protein and tested up to 15 minutes exposure to ECL.

Thanks

Read More

Abcam community

Verified customer

Asked on Nov 23 2012

Answer

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

I would like to reassure you that this antibody is tested and covered by our 6 month guarantee for WB and human samples. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.

I would be pleased to help troubleshoot, and also obtain more information for our quality monitoring records. In order to proceed with this, I have enclosed a technical questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily which will enable us to better understand the problem and provide assistance.

I would appreciate if you are also able to provide an image which would help us to assess the results.

Thank you for your time and cooperation. We look forward to receiving the completed questionaire.

Order Details
Antibody code:

Problem
Choose: Non-specific band Multiple bands No signal or weak signal High background

Lot number

Purchase order number
or preferably Abcam order number:



General Information
Antibody storage conditions (temperature/reconstitution etc)


Description of the problem (high background, wrong band size, more bands, no band etc.)


Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.)


Sample preparation (Buffer/Protease inhibitors/Heating sample etc.)


Amount of protein loaded


Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.)


Transfer and blocking conditions (Buffer/time period, Blocking agent etc.)


Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Detection method (ECL, ECLPlus etc.)


Positive and negative controls used (please specify)



Optimization attempts (problem solving)
How many times have you tried the Western?



Have you run a "No Primary" control?
Yes No

Do you obtain the same results every time?
Yes No
e.g. are the background bands always in the same place?


What steps have you altered?


Additional Notes:


Image:
We would appreciate if you are able to provide an image (including molecular weight markers) which would help us to asess the results.

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回复于 Nov 23 2012

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