Overview

  • Product name

    Anti-VCAM1 antibody [EPR5047] - Low endotoxin, Azide free
    See all VCAM1 primary antibodies
  • Description

    Rabbit monoclonal [EPR5047] to VCAM1 - Low endotoxin, Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, IHC-P, Flow Cyt, IHC-Fr, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Mouse VCAM1.
    (Peptide available as ab156177)

  • Positive control

    • Human fetal liver, HuT 78, NIH 3T3, Mouse brain, Mouse kidney, Mouse spleen, Rat brain, Rat kidney and Rat spleen lysates. IHC-P: Human and Mouse spleen FFPE tissue sections.
  • General notes

    ab215380 is a carrier-free antibody designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

    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.

Applications

Our Abpromise guarantee covers the use of ab215380 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 Use at an assay dependent concentration. Detects a band of approximately 100 kDa (predicted molecular weight: 81 kDa).Can be blocked with VCAM1 peptide (ab156177).
IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocol.

Flow Cyt Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.

Target

  • Function

    Important in cell-cell recognition. Appears to function in leukocyte-endothelial cell adhesion. Interacts with the beta-1 integrin VLA4 on leukocytes, and mediates both adhesion and signal transduction. The VCAM1/VLA4 interaction may play a pathophysiologic role both in immune responses and in leukocyte emigration to sites of inflammation.
  • Tissue specificity

    Expressed on inflammed vascular endothelium, as well as on macrophage-like and dendritic cell types in both normal and inflammed tissue.
  • Sequence similarities

    Contains 7 Ig-like C2-type (immunoglobulin-like) domains.
  • Domain

    Either the first or the fourth Ig-like C2-type domain is required for VLA4-dependent cell adhesion.
  • Post-translational
    modifications

    Sialoglycoprotein.
  • Cellular localization

    Membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • CD106 antibody
    • CD106 Antigen antibody
    • INCAM 100 antibody
    • INCAM-100 antibody
    • L1CAM antibody
    • MGC99561 antibody
    • V-CAM 1 antibody
    • Vascular Cell Adhesion Molecule 1 antibody
    • Vascular cell adhesion protein 1 antibody
    • VCAM 1 antibody
    • VCAM-1 antibody
    • VCAM1 antibody
    • VCAM1_HUMAN antibody
    see all

Images

  • Flow Cytometry analysis of K562 (human chronic myelogenous leukemia) cells labeling VCAM1 with purified ab134047 at 1/40 dilution (10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor®488) at 1/2000 dilution was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134047).

  • VCAM1 was immunoprecipitated from Human fetal liver lysate with ab134047 at 1/110 dilution.

    Western blot was performed from the immunoprecipitate using ab134047 at 1/1000 dilution.

    VeriBlot for IP secondary antibody (Peroxidase conjugated),was used as secondary antibody at 1/1000 dilution.

    Lane 1:Human fetal liver lysate

    Blocking and dilution buffer:5% NFDM/TBST

    Exposure time: 1 second

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134047).

  • Immunofluorescent staining of K562 cells (fixed in 4% PFA, permeabilized with 0.1% Triton X 100) using purified ab134047 at a dilution of 1/250. An Alexa Fluor® 488 goat anti-rabbit antibody was used as the secondary at a dilution of 1/500 and the cells were counter stained with DAPI. The negative control is shown in the bottom right hand panel - for the negative control, Alex Fluor® 594 goat anti-mouse was used at a dilution of 1/500.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134047).

  • IHC image of VCAM1 staining in Mouse spleen formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with unpurified ab134047, 1/200 dilution, for 15 mins at room temperature. A goat anti-rabbit biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134047).

  • This IHC data was generated using the same anti-VCAM1 antibody clone, EPR5047, in a different buffer formulation (cat# ab134047).

    IHC image of VCAM1 staining in Human spleen formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with unpurified ab134047, 1/200 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • This IHC data was generated using the same anti-VCAM1 antibody clone, EPR5047, in a different buffer formulation (cat# ab134047).

    Immunohistochemical staining of paraffin embedded human tonsil with purified ab134047 at a dilution of 1/500. A pre-diluted HRP polymer for rabbit/mouse IgG was used as the secondary antibody and the sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

References

This product has been referenced in:

  • Wu F  et al. CXCR2 is essential for cerebral endothelial activation and leukocyte recruitment during neuroinflammation. J Neuroinflammation 12:98 (2015). WB ; Mouse . Read more (PubMed: 25990934) »
  • Coon BG  et al. Intramembrane binding of VE-cadherin to VEGFR2 and VEGFR3 assembles the endothelial mechanosensory complex. J Cell Biol 208:975-86 (2015). Read more (PubMed: 25800053) »
See all 6 Publications for this product

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