Overview

  • Product name
    Anti-ICAM1 antibody [YN1/1.7.4]
    See all ICAM1 primary antibodies
  • Description
    Rat monoclonal [YN1/1.7.4] to ICAM1
  • Host species
    Rat
  • Tested applications
    Suitable for: IP, IHC-Fr, Blocking, WB, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse
  • Immunogen

    The details of the immunogen for this antibody are not available.

  • Positive control
    • PMA or LPS stimulated T-cell line such as EL-4 or 18-81

Properties

Applications

Our Abpromise guarantee covers the use of ab25375 in the following tested applications.

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

Application Abreviews Notes
IP Use at an assay dependent concentration. PubMed: 2573511
IHC-Fr Use at an assay dependent concentration. PubMed: 8943409
Blocking Use at an assay dependent concentration. PubMed: 2981916
WB Use at an assay dependent concentration. Detects a band of approximately 95 kDa (predicted molecular weight: 85-110 kDa). PubMed: 8095510
Flow Cyt Use at an assay dependent concentration.

Target

  • Function
    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.
  • Sequence similarities
    Belongs to the immunoglobulin superfamily. ICAM family.
    Contains 5 Ig-like C2-type (immunoglobulin-like) domains.
  • Post-translational
    modifications
    Monoubiquitinated, which is promoted by MARCH9 and leads to endocytosis.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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

Images

  • Flow Cytometry analysis of BALB/c mouse splenocytes labeling ICAM1 with ab25375 at 0.3 μg/106 cells (purple). A Mouse Anti-Rat IgG2b-CY5 was used as the secondary antibody. Grey - Isotype Control, Rat IgG2b-UNLB, followed by Mouse Anti-Rat IgG2b-CY5.

References

This product has been referenced in:
See all 19 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Pulmonary Microvascular Endothelial Cells)
Permeabilization
Yes - 0.5% Triton in PBS 10 min on ice
Specification
Pulmonary Microvascular Endothelial Cells
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 25°C
Fixative
Methanol:Acetone

Jose Vazquez-Medina

Verified customer

Submitted Nov 15 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate
Sample
Mouse Tissue sections (Brain)
Specification
Brain
Permeabilization
Yes - 0.1% Triton
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Mar 13 2015

Question
Answer

Thank you for your call today and for your interest in these antibodies.

Since we do not have a CD54 antibody that hasalready been tested in IHC-P with mouse samples, I can offer a discount off a future purchase if you'd liketest a CD54 antibodynow, test it in IHC-P, and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be worth one free primary antibody on a future purchase.

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and test a CD54 antibody in IHC-P. I will then send a discount code.

2. Purchasethe CD54 antibodyeither by phone, fax, or online (www.abcam.com).

3. Test it in IHC-P.

4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for anyprimary antibodyordered and the discount code is valid for 4 months after issue.
Please remember that submission of the Abreview is sufficient for the discount code to become active.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if the CD54 antibody turns out to be unsuitable for IHC-P, you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

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Question
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 product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement of ab25375.

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 us know.

I wish you the best of luck with your research.

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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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Answer

Perdona por la tardanza en contestar, estaba chequeando con mis compañeros del laboratorio las distintas opciones de reactividad cruzada de este anticuerpo con conejo. Desgraciadamente no hay muchos datos al respecto, ya que ninguno de ellos se ha testado en esta especie, y muchos de los epítopos no han sido mapeados, por lo que no puede llevarse a cabo un BLAST.

Así pues, si sigues interesada en testarlo en conejo, a pesar de no tener datos concretos acerca de las posibilidades de reactividad, te puedo mandar un código para canjear por un anticuerpo gratuito si nos envías los resultados (independientemente de si estos resultan positivos o no).

Para detectar I CAM1 yo me inclinaría por ab25375 o ab27294, y en el caso de V CAM1 por ab78712 o ab98954.

Es importante que antes de realizar el pedido te emita el código valido para el anticuerpo gratuito por participar en el sistema Abreviews, así que te pediría que antes de hacer el pedido, por favor, esperes a que te lo comunique.

¿Qué tal han ido los resultados con ab25072?

Respecto a la duda de los Abreviews, te cuento. Si se trata de una aplicación o especie que no se ha testado hasta el momento, la compensación por participar en el sistema Abreview es un código (emitido ANTES de realizar el pedido) válido por un anticuerpo primario completamente gratuito, y también una serie de puntos (Abpoints) canjeables por distintas opciones.

Si, por el contrario, los resultados enviados corresponden a información adicional sobre especies o aplicaciones ya testadas, en ese caso se ofrecen (Abpoints), pues ayudan a completar la información y referencias del anticuerpo.

Respecto a los anticuerpos que ya habéis probado en el pasado, siempre podéis publicar los resultados, pero aunque sean nuevas aplicaciones / especies no podremos ofreceros un código de descuento ya que el pedido se ha realizado con anterioridad a dicho código. Aun así, podéis mandar los resultados, y ser compensados con puntos.

Espero haberte aclarado. Quedo a la espera de recibir tu respuesta para que me indiques como proceder. Si tienes más dudas, por favor, no dudes en volvernos a contactar.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain)
Specification
Brain
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C

Dr. Thomas Weissensteiner

Verified customer

Submitted Oct 22 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain)
Specification
Brain
Fixative
PLP
Permeabilization
Yes - 0.05% Tween-20 washes
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Aug 19 2009

Answer

Thank you for your enquiry. I have enquired at the originator of the product but unfortunately, they do not have the information that you are requesting. I am sorry I can't be of more assistance in this occasion. If there is anything else I can help you with, please do not hesitate to contact me.

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Question

Here is my reply to your questions. Batch number: lot # 155779 Purchase order number 61281D or preferably Abcam order number: General Information Antibody storage conditions (temperature/reconstitution etc): 4 C Description of the problem (high background, wrong band size, more bands, no band etc.): Not picking up anything definitive b/t 85 and 110 kDa Sample (Species/Cell extract/Nuclear extract/Purified protein/Recombinant protein etc.): Mouse Brain Whole Cell Lysates Sample preparation (Buffer/Protease inhibitors/Heating sample etc.): 1X 'Cell Signaling' Cell Lysis Buffer Amount of protein loaded: Loading max amount of 350ug Electrophoresis/Gel conditions (Reducing or Non-reducing gel, % of the gel etc.): Reducing, 10% gel Transfer and blocking conditions (Buffer/time period, Blocking agent etc.): Transfer 100V, 30 min. Block 5%milk for 1 hour room temp. Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step): Ab25375, 1:1000 dilution overnight @ 4C Wash 5X 15 min each Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step): Ab6734-1 Detection method (ECL, ECLPlus etc.): Amersham ECL Positive and negative controls used (please specify): Using Hela cells, max loading of 30ul, boiled for 5 min. Optimization attempts (problem solving): Changed dilution of secondary from 1:1000 to 1:500, max loading the protein to pick up the band, Re- running the blots, etc. How many times have you tried the Western? 10 times Have you run a "No Primary" control? Yes Do you obtain the same results every time? Yes What steps have you altered? See optimization attempts above Additional Notes: We are having a difficult time even picking up the pos. Hela control. We have used 5 different lots of Hela controls and none show up with this antibody. We are getting many non-specific bands. I think the antibody might be unstable, since from the first time we have used it, we have never had a positive result.

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Answer

Thank you for providing these details regarding your protocol. Ab25375 should detect a band at approximately 58 kDa; did you see a band at this size? This particular antibody has only so far been tested for cross-reactivity with mouse and has not yet been tested for cross-reactivity with human. So even though HeLa samples should in theory be a good control (as CD54 is expressed on endothelial cells) we don't know if this antibody detects human ICAM1. To help decrease the amount of non-specific bands that you are seeing, I would suggest loading less lysate on your gel (we recommend 20-30 ug; 350 ug is too much) and decrease the amount of primary and secondary that you are using as well as the incubation periods. You can try the primary at 1:1000 for 2 hrs at RT or at 1:2000. For the secondary you can try it at a dilution of 1:5000. Please let me know if these suggestions help. If not, I can certainly offer you a free of charge replacement, a credit note or a refund.

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