Anti-ICAM1 抗体 [MEM-111] (ab2213)

Reviews (12)Q&A (23)References (29)

製品の概要

  • 製品名

    Anti-ICAM1 antibody [MEM-111]
    ICAM1 一次抗体 製品一覧

  • 製品の詳細

    Mouse monoclonal [MEM-111] to ICAM1

  • 由来種

    Mouse

  • 特異性

    This antibody reacts with human CD54 antigen.

  • アプリケーション

    適用あり: ICC/IF, ELISA, WB, IHC-P, IHC-Fr, Flow Cyt, Sandwich ELISAmore details

  • 種交差性

    交差種: Cow, Human

  • 免疫原

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

  • ポジティブ･コントロール

    • 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.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

ターゲット情報

  • 機能

    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.

  • 配列類似性

    Belongs to the immunoglobulin superfamily. ICAM family.
    Contains 5 Ig-like C2-type (immunoglobulin-like) domains.

  • 翻訳後修飾

    Monoubiquitinated, which is promoted by MARCH9 and leads to endocytosis.

  • 細胞内局在

    Membrane.
  • Information by UniProt

  • 参照データベース

    • 別名

      • 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) 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 antibody
      • Surface antigen of activated B cells, BB2 antibody
      see all

    画像

    • Immunocytochemistry/ Immunofluorescence - Anti-ICAM1 antibody [MEM-111] (ab2213)
      Immunocytochemistry/ Immunofluorescence - Anti-ICAM1 antibody [MEM-111] (ab2213)Ross, S.L. et al PLoS One. 2017 Aug 24;12(8):e0183390. doi: 10.1371/journal.pone.0183390. eCollection 2017 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

      Representative images demonstrating ICAM-1 and FAS induction by recombinant cytokines and BiTE®-activated T cells.

      Immunofluorescence images showing ICAM-1 upregulation in NUGC4 cells by (A) 12.5ng/ml each IFNγ + TNFα or (B) 33 pM BiTE® (A, B, blue = nuclear stain, red = ICAM-1 staining). Representative images of SW620 cells showing upregulation of ICAM-1 by (C) 12.5ng/ml each IFNγ + TNFα or (D) BiTE®-activated T cells.  Scale bar = 30 μm.

      ICAM1 was detected with ab2213.

      (After Figure S5 of Ross et al).

    • Western blot - Anti-ICAM1 antibody [MEM-111] (ab2213)
      Western blot - Anti-ICAM1 antibody [MEM-111] (ab2213)
      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.

    • Flow Cytometry - Anti-ICAM1 antibody [MEM-111] (ab2213)
      Flow Cytometry - Anti-ICAM1 antibody [MEM-111] (ab2213)This image is courtesy of an Abreview submitted by Dr Karppiah Muthumani

      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

    • Sandwich ELISA - Anti-ICAM1 antibody [MEM-111] (ab2213)
      Sandwich ELISA - Anti-ICAM1 antibody [MEM-111] (ab2213)
      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
    • Western blot - Anti-ICAM1 antibody [MEM-111] (ab2213)
      Western blot - Anti-ICAM1 antibody [MEM-111] (ab2213)This image is courtesy of an anonymous 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

    参考文献

    This product has been referenced in:

    • Zhang K  et al. Activation of brain endothelium by Escherichia coli K1 virulence factor cglD promotes polymorphonuclear leukocyte transendothelial migration. Med Microbiol Immunol 208:59-68 (2019). Read more (PubMed: 30171337) »
    • Jiao H  et al. The Ovotransferrin-Derived Peptide IRW Attenuates Lipopolysaccharide-Induced Inflammatory Responses. Biomed Res Int 2019:8676410 (2019). Read more (PubMed: 30719449) »
    See all 29 Publications for this product

    ab2213 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    レビューと Q&A

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    レビューを送る 質問を送る

    1-10 of 35 Abreviews or Q&A

    Blocking abreview for Anti-ICAM1 antibody [MEM-111]

     Excellent
    Abreviews
    Application
    Blocking
    Sample
    Streptococcus pneumoniae Cell (Primary Bronchial Epithelial cells)
    Specification
    Primary Bronchial Epithelial cells
    Read More

    Dr. Selvarani Vimalanathan

    Verified customer

    投稿 Aug 03 2018

    Immunocytochemistry abreview for Anti-ICAM1 antibody [MEM-111]

     Good
    Abreviews
    abreview image
    Application
    Immunocytochemistry
    Sample
    Human Cultured Cells (BEAS-2B Epithelial cells)
    Permeabilization
    Yes - 0.5% saponin
    Specification
    BEAS-2B Epithelial cells
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C
    Fixative
    Formaldehyde
    Read More

    Dr. Selvarani Vimalanathan

    Verified customer

    投稿 Aug 04 2017

    Western blot abreview for Anti-ICAM1 antibody [MEM-111]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Cancer cell lines total lysate)
    Gel Running Conditions
    Reduced Denaturing (10%)
    Loading amount
    20 µg
    Specification
    Cancer cell lines total lysate
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    投稿 Dec 07 2015

    Western blot abreview for Anti-ICAM1 antibody [MEM-111]

     Inconclusive
    Abreviews
    Application
    Western blot
    Loading amount
    40 µg
    Gel Running Conditions
    Non-reduced Non-Denaturing (Native)
    Sample
    Rhesus monkey Cell lysate - whole cell (RF/6A cells from ATCC)
    Specification
    RF/6A cells from ATCC
    Treatment
    5.5 mM glucose, 30 mM glucose, 30 mM glucose + 0.1 mM BAPN for 10 days
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    投稿 May 26 2014

    Question

    I hope this email finds you well. I was just writing to you with regards to an order I recently placed. This was for an anti-ICAM1 antibody [MEM-111] (catalogue number ab2213). On the datasheet provided and on the vial it informs me that there should be 100ug at 1mg/ml, however when aliquoting the antibody I only had 90ul instead of 100ul, so had 10ug less. I thought you should know this to avoid any further issues in the future when sending out antibodies. I have been buying all my antibodies from you and this has been an isolated issue so perhaps it is a product specific problem with this item, but I thought I should raise the issue with you.
    Kind regards,

    Read More
    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.

    Read More

    Immunohistochemistry (Frozen sections) abreview for Anti-ICAM1 antibody [MEM-111]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Rat Tissue sections (rat cortex (injured))
    Specification
    rat cortex (injured)
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.1% triton X-100 in blocking buffer
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    投稿 Mar 20 2013

    Question

    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.
    Please find attached

    Thanks,

    Read More
    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.

    Read More

    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.

    Read More
    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.

    Read More

    Question

    Dear,
    I ordered the following antibodies and received them at the end of September:
    - Ab54426 (anti-CD62P)
    - Ab91353 (Mouse IgG1 – isotype control)
    - Ab 18981 (anti-CD62E)
    - Ab27478 (rabbit polyclonal IgG)
    - Ab2213 (anti-ICAM1)

    I stored them at +4oC till today, but I have noticed today that your storage instructions recommend to aliquot and store most of the antibodies at -20oC or -80oC.
    Could this have had an effect on my stainings?
    Kind regards,

    Read More
    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.

    Read More

    Question

    Phone call reporting difficulty in using the Anti-ICAM1 antibody [MEM-111] (ab2213) in ICC.

    Read More
    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.

    Read More

    1-10 of 35 Abreviews or Q&A

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