概述

  • 产品名称
    Anti-ICAM1抗体[MEM-111]
    参阅全部 ICAM1 一抗
  • 描述
    小鼠单克隆抗体[MEM-111] to ICAM1
  • 宿主
    Mouse
  • 特异性
    This antibody reacts with human CD54 antigen.
  • 经测试应用
    适用于: ICC/IF, ELISA, WB, IHC-P, IHC-Fr, Flow Cyt, Sandwich ELISAmore details
  • 种属反应性
    与反应: Cow, Human
  • 免疫原

    Tissue/ cell preparation (Human). (Burkitt's lymphoma cell line RAJI).

  • 阳性对照
    • For WB it is highly recommended to use TNF-alpha activated HUVEC cells as a positive control. Sandwich ELISA: supernatant of HUVEC cell line. Indirect ELISA: endothelial cell line derived from bovine pulmonary artery. RAJI cell line,K562 leukemia cell line, JY cell line, Activated T-lymphocytes, Thymus, RE cells.

性能

应用

Our Abpromise guarantee covers the use of ab2213 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
ICC/IF 1/100.
ELISA Use a concentration of 25 µg/ml. Use at a dilution of 1/1000 for Indirect ELISA, 1hr, RT. The antibody reacts with tissue - endothelial cell line derived from bovine pulmonary artery.
WB Use at an assay dependent concentration. Use under non-reducing conditions.
IHC-P Use a concentration of 10 µg/ml.
IHC-Fr Use at an assay dependent concentration.
Flow Cyt Use a concentration of 2 µg/ml.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

Sandwich ELISA Use a concentration of 5 µg/ml. Can be paired for Sandwich ELISA with Rabbit polyclonal to ICAM1 (Biotin) (ab7815).

For sandwich ELISA, use this antibody as Capture at 5µg/ml with ab7815 as Detection.

靶标

  • 功能
    ICAM proteins are ligands for the leukocyte adhesion protein LFA-1 (integrin alpha-L/beta-2). During leukocyte trans-endothelial migration, ICAM1 engagement promotes the assembly of endothelial apical cups through ARHGEF26/SGEF and RHOG activation. In case of rhinovirus infection acts as a cellular receptor for the virus.
  • 序列相似性
    Belongs to the immunoglobulin superfamily. ICAM family.
    Contains 5 Ig-like C2-type (immunoglobulin-like) domains.
  • 翻译后修饰
    Monoubiquitinated, which is promoted by MARCH9 and leads to endocytosis.
  • 细胞定位
    Membrane.
  • Information by UniProt
  • 数据库链接
  • 别名
    • Antigen identified by monoclonal antibody BB2 antibody
    • BB 2 antibody
    • BB2 antibody
    • CD 54 antibody
    • CD_antigen=CD54 antibody
    • CD54 antibody
    • Cell surface glycoprotein P3.58 antibody
    • Human rhinovirus receptor antibody
    • ICAM 1 antibody
    • ICAM-1 antibody
    • ICAM1 antibody
    • ICAM1_HUMAN antibody
    • intercellular adhesion molecule 1 (CD54), human rhinovirus receptor antibody
    • Intercellular adhesion molecule 1 antibody
    • Major group rhinovirus receptor antibody
    • MALA 2 antibody
    • MALA2 antibody
    • MyD 10 antibody
    • MyD10 antibody
    • P3.58 antibody
    • Surface antigen of activated B cells, BB2 antibody
    see all

图片

  • All lanes : Anti-ICAM1 antibody [MEM-111] (ab2213)

    Lane 1 : TNF-alpha activated HUVEC cells
    Lane 2 : TNF-alpha nonactivated HUVEC cells

    Performed under non-reducing conditions.

    Observed band size: 90 kDa
    why is the actual band size different from the predicted?



    Lower bands represent tubulin as a loading control.

  • Standard Curve for ICAM1 (Analyte: ab82125) dilution range 1pg/ml to 1ug/ml using Capture Antibody Mouse monoclonal [MEM-111] to ICAM1 (ab2213) at 5ug/ml and Detector Antibody Rabbit polyclonal to ICAM1 (Biotin) (ab7815) at 0.5ug/ml
  • ab2213 staining human PBL cells by flow cytometry.  Cells were formaldehyde fixed and gated on B-cells.  The primary antibody was incubated with the sample for 30 minutes at 25°C.  The secondary antibody was ab6810 (a FITC conjugated chicken polyclonal to mouse IgG - H&L).  Both antibodies were used undiluted.

    See Abreview

  • All lanes : Anti-ICAM1 antibody [MEM-111] (ab2213) at 1/3000 dilution

    Lane 1 : Western Markers.
    Lane 2 : Whole cell lysate prepared from human HUVEC cells (untreated) at 100,000 cells.
    Lane 3 : Whole cell lysate prepared from human HUVEC cells (treated with 1 ng/ml TNF-alpha for 24 h) at 100,000 cells.

    Secondary
    All lanes : Rabbit Anti-Mouse IgG H&L (HRP) (ab6728)

    Observed band size: 95 kDa why is the actual band size different from the predicted?
    Additional bands at: 40 kDa (possible non-specific binding)

    See Abreview

文献

This product has been referenced in:
  • Li Y  et al. Early intervention with mesenchymal stem cells prevents nephropathy in diabetic rats by ameliorating the inflammatory microenvironment. Int J Mol Med 41:2629-2639 (2018). Read more (PubMed: 29484379) »
  • Kotha S  et al. Microvasculature-directed thrombopoiesis in a 3D in vitro marrow microenvironment. PLoS One 13:e0195082 (2018). ICC/IF . Read more (PubMed: 29617409) »
See all 23 Publications for this product

客户评价及客户问答

1-10 of 23 Q&A

Answer

I sorry to hear about the volume of the antibody; we always fill vials 4-5ul extra so quick spin is required to extract the whole volume. I am sure you have given a quick spin to vial before opening it, if it hasn't been done then it will worth to try.

On the other hand I have let our logistic team know about this issue and I can reassure you that they have taken the necessary actions.

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Answer

Thank you for your enquiry.

As the Swiss-Prot database highlights, the mature ICAM1 protein is heavily N-Glycosylated. There at least 9 potential site for this type of post-translational modification which can significantly alter the band size. I would advise you to consult these sites for further information:

http://www.uniprot.org/uniprot/P05362

http://www.uniprot.org/uniprot/Q00238

If you need any further assistance in the future, please do not hesitate to contact me.

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Question

Thank you very much,

We used ICAM-1 (Ab, ab2213), but this antibody gives many unspecific bands. I tried it in many different conditions, however, it still gives me the same results.

· Lysis buffer - would be very important to know

We are using special a kit from abcam ab65400 (Plasma membrane protein extraction kit) in which lysis buffer is included.

· Amount of protein loaded - (protein endogenously expressed or transfected cells)

20-30 μg protein per lane.

· Electrophoresis and transfer conditions

The samples were run at 100 and 150 V, one dimensional SDS-PAGE gel was used. For transfering, Protein transfer verifiedusing Ponceau red.

· Blocking conditions

We used 5% and 3% fat free dry milk and also we tried 5% and 3% BSA. Blocking time, once 1 hour and in other run was 30 minutes at 25˚C

· Primary antibody conditions (diluent/dilution, incubation time/temperature)

Concentration or dilution

1/1000 to 1/250 under reducing and non-reducing condition.

· Diluent buffer

1X PBS+0.1% Tween in 5% BSA.

· Incubation time

Once over night at 4C and another 1 hour and 30 minutes at room temperature.

· Secondary antibody conditions (diluent/dilution, incubation time/temperature)

Secondary antibody species gout which reacts against mouse antibodies, the dilution is from 1/10000 to 1/20000 in 1X PBS+0.1% Tween in 5% BSA for one hour at 25˚C.

Wash steps and washing buffer: 1X PBS+0.1% Tween 3 times each one 10 minutes.

· Detection method:

ECL (super signal west pico chemiluminescent)

· Positive control
I used Calnexin, and Na/K ATPase (ab7671) as membrane markerThank you for your understanding and co-operation in this matter. I look forward to hearing from you soon and resolving this issue as soon as possible.

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Answer

Thank you for your response.

Could you please specify the size of the non-specific bands? Are they lower or higher than the expected size (58 kDa)? Could you attach an image (saved as a jpeg file) to your e-mail so that I can have a look at the bands.

I look forward to hearing from you soon.

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Answer

Thank you for your enquiry.

I can confirm that ab18414 can be stored at 4oC as described on the datasheet. This should not be aliquotted and frozen.

The others I would recommend to aliquot and freeze immediately. Keeping them at 4oC for a few weeks should not significantly affect the staining. However, we would recommend to follow the individual storage instructions provided on the individual datasheets as soon as possible after delivery. This will help to ensure that the antibody remains stable and that we can continue to provide our guarantee.

I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

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Question
Answer

Thank you for calling us yesterday and for alerting us to the problem you are experiencing with our product. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

As we discussed on the phone, I have attached our questionnaire so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results.

This information will also allow us to investigate this case internally and initiate additional testing where necessary.

I look forward to receiving your reply.

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Answer

I am very pleased to hear you would like to accept our offer and test ab54426 in IHC-P.

DISCOUNT CODE: ######

Expiration date: ######

This code will give you: 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for ab54426 used in IHC-P and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.

For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways:

1) Call to place your order and mention the code to our customer service department;
2) Include the code in your fax order;
3) Place your order on the web and enter the promotional code.

With regards to the isotype controls:
For ab2213, the light chain type is unknown, so I would recommend to use a general IgG2a isotype control rather than a specific IgG kappa chain isotype control. For example the following product:

ab91361 Mouse IgG2a [ICIGG2A] - Isotype Control
https://www.abcam.com/index.html?datasheet=91361 https://www.abcam.com/index.html?datasheet=91361.

However, please note this is not tested in IHC-P. Unfortunately, I am sorry we do not have any suitable isotype controls tested in IHC-P in this case.

ab27478 is a suitable isotype control for ab18981
ab91353 is a suitable isotype control for ab54426

Any feedback that you can provide regarding ab54426 will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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Question
Answer

Thank you for contacting us.

I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.

Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

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Answer

Thank you for contacting Abcam regarding ab2213.

I am sorry that you have been experiencing difficulities with this antibody in IHC. Based on the information you provided I agree that this product is not performing as stated on the datasheet. I am happy to offer a replacement or credit per our Abpromise guarantee. Please let me know which you would prefer. Can you please include your order number in your response?

I look forwrad to your reply so that I may assist you further. Please do not hesitate to contact me if you have any additional questions.

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Answer

Thank you for the response.

Any monoclonal antibody is produced by a single immune cell, which has been cloned. These will only recognize a single epitope of the target protein. This epitope typically is 5-10 amino acids in length. Therefore any single monoclonal antibody will only recognize a region within a single domain and not stain both the intracellular and extracellular region of the target protein.

Polyclonal antibodies are created by many different immune cells and will contain multiple epitope recognition sites. Although the amount of antibody specific to a particular epitope will vary from lot-to-lot, it is possible for a polyclonal to therefore contain antibodies which will recognize different domains.

I have reviewed the antibodies which you have listed as well as other polyclonals in our catalog to try and find a product which might be able to recognize the intracellular and extracellular regions.

ab2213Anti-ICAM1 antibody [MEM-111]: This monoclonal antibody was created with an immunogen from a whole tissue / cell preparation (Human). We have not mapped the exact location of the epitope. Therefore the epitope recognized might be intracellular, transmembrane or extracellular.

ab25375Anti-ICAM1 antibody [YN1/1.7.4]: This product is also monoclonal.The details of the immunogen for this antibody are not available. However as a monoclonal, this product will not be able to recognize sites in multiple domains.

ab7815Anti-ICAM1 antibody (Biotin): This product is a rabbit polyclonal and has been raised against the whole protein. As such it is possible that the product will contain antibodies against both extracellular and intracellular regions. We do not test for this type of response and cannot guarantee that the productwill recognizeboth domains. Please be aware that this product is also biotin labeled.

Our other rabbit polyclonal antibodies were each created from recombinant fragments and would also not be able to recognize both extracellular and intracellular domains.

I hope this information is helpful. Please do not hesitate to contact me with further questions.

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Question
Answer

Thank you for your reply.

The options of resolving this case are to replace the antibody free of charge with another antibody (different lot; same target; different target)or to issue a credit or refund.

I assume from the email conversation with my colleague that you have used human tissue?

We carry several ICAM1-antibodies that have been tested in human and IHC-P and from which you can choose from:
ab53013, ab67217, ab109361, ab20144, ab67345and ab89378.

I look forward to hear back from you and resolve this case.

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1-10 of 23 Q&A

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

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