Recombinant
RabMAb

Recombinant Anti-ICAM1 antibody [EPR22161-284] (ab222736)

Overview

  • Product name

    Anti-ICAM1 antibody [EPR22161-284]
    See all ICAM1 primary antibodies
  • Description

    Rabbit monoclonal [EPR22161-284] to ICAM1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, ICC/IF, IPmore details
  • Species reactivity

    Reacts with: Mouse
  • Immunogen

    Recombinant fragment within Human ICAM1 aa 1-500. The exact sequence is proprietary.
    Database link: P13597

  • Positive control

    • WB: Mouse kidney, spleen and liver tissue lysate; A20 whole cell lysate. IHC-P: Mouse lung and spleen tissue. ICC/IF: A20 cells. Flow: Mouse primary splenocytes. IP: Mouse spleen lysate.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab222736 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
WB 1/1000. Detects a band of approximately 110 kDa (predicted molecular weight: 57 kDa).
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Flow Cyt 1/500.
ICC/IF 1/50.
IP 1/30.

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) 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

Images

  • All lanes : Anti-ICAM1 antibody [EPR22161-284] (ab222736) at 1/1000 dilution

    Lane 1 : Mouse kidney tissue lysate
    Lane 2 : Mouse spleen tissue lysate
    Lane 3 : Mouse liver tissue lysate
    Lane 4 : A20 (mouse reticulum sarcoma b lymphocyte) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 57 kDa
    Observed band size: 110 kDa
    why is the actual band size different from the predicted?



    Blocking/Diluting buffer and concentration: 5% NFDM/TBST.

    Exposure time: 48 seconds.

    The 110-KDa apparent molecular mass is due to glycosylation (PMID: 1680698).

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A20 (mouse reticulum sarcoma cell line) cells labeling ICAM with ab222736 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in A20 cells with distinct staining of the target protein being transported from Golgi to cell surfaces (PMID: 17486117) is observed. Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) was used as a countertsain (Red) at 1/200 dilution. The nuclear counterstain is DAPI (Blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Alexa Fluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution.

  • Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling ICAM1 using ab222736 at 1/1000 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on the marginal zone of mouse spleen (PMID: 12130787; PMID: 17947659) is observed. Counterstained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) ready to use.

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    Immunostaining was performed on a Leica Biosystems BOND® RX instrument. The section was incubated with ab222736 for 30 mins at room temperature.

  • ICAM1 was immunoprecipitated from 0.35 mg Mouse spleen lysate with ab222736 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab222736 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/5000 dilution.

    Lane 1: Mouse spleen tissue lysate 10 μg (input).
    Lane 2: ab222736 IP in mouse spleen tissue lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab222736 in Mouse spleen lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 3 minutes.

    110-kDa molecular mass is due to glycosylation (PMID: 1680698).

  • Flow cytometric analysis of Mouse primary splenocytes labeling ICAM1 with ab222736 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabeled control (Cell without incubation with primary antibody and secondary antibody, Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/200 dilution was used as the secondary antibody. Gated on viable cells.

    ICAM1 negative population (left half of red peak) was also observed in mouse splenocytes which is consistent with IHC staining pattern.

  • Immunohistochemical analysis of paraffin-embedded mouse lung tissue labeling ICAM1 using ab222736 at 1/1000 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Membranous and cytoplasmic staining on mouse lung (PMID: 14568945; PMID: 9802985) is observed. Counterstained with DAPI.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) ready to use.

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    Immunostaining was performed on a Leica Biosystems BOND® RX instrument. The section was incubated with ab222736 for 30 mins at room temperature.

References

ab222736 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab222736.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up