Overview

  • Product name
    Anti-VCAM1 antibody [M/K-2] (Biotin)
    See all VCAM1 primary antibodies
  • Description
    Rat monoclonal [M/K-2] to VCAM1 (Biotin)
  • Host species
    Rat
  • Conjugation
    Biotin
  • Tested applications
    Suitable for: Blocking, Inhibition Assay, Flow Cyt, IP, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse
  • Immunogen

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

Applications

Our Abpromise guarantee covers the use of ab24942 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
Blocking Use at an assay dependent concentration.

Blocks lymphocyte adhesion in Whitlock-Witte long-term bone marrow cultures.

Inhibition Assay Use at an assay dependent concentration.

Inhibition of lymphopoiesis in Whitlock-Witte cultures.

Flow Cyt Use 1µg for 106 cells.

ab18403 - Rat monoclonal IgG1, is suitable for use as an isotype control with this antibody.

IP Use at an assay dependent concentration.

Immunoprecipitates a peptide that gives a single band on SDS PAGE gels with an apparent Mr of ~100 kDa under reducing conditions and 92 kDa under non reducing conditions.

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

References

ab24942 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Question

Inquiry: Hi, I have tested this antibody a couple of times using an ICC protocol for mouse aortic endothelial cells that were stimulated with TNF alpha to express VCAM1. The staining I am getting is very weak and I just wondered if you have any suggestions for a protocol to follow and the best concentration of antibody to use. My protocol was as follows Remove culture medium from each well and wash twice with PBS Add 1ml 4% paraformaldehyde solution in PBS for 15-20mins at room temperature Wash cells twice with PBS and then cover with 1ml wash buffer ( 0.1% BSA in PBS)(Coverslips can be stored in fridge for up to 3 months or stained immediately) Wash the coverslips containing the fixed cells two times with 1ml wash buffer. Blocking step – incubate cells for 45mins with either 1% BSA in PBS or 10% serum from which secondary antibody is raised. 1 Incubate cells with primary antibody tried 1:100, 1:200 and 1:500 very weak staining at 1:100 and 1:200 but not much at 1:500 (diluted in 1% BSA) in humidified chamber overnight at 4⁰C Decant the solution and wash the cells 3 times with PBS 0.1% BSA Incubate cells with fluorescent secondary antibody Streptavidin alexa fluor 488 1:200 for 1 hour at room temperature in the dark. Decant secondary antibody solution and wash with PBS 3 x 5 mins in the dark. Incubate cells with DAPI for 1 min Rinse with PBS 4 x 5 min washes Remove the coverslips from wells and blot to remove any water, dispense 1 drop of anti-fade mounting medium onto microscope slide and then mount coverslip with cells facing towards the microscope slide. Either store at -20C or visualise under fluorescent microscope. I also wondered if this antibody has been tested in paraffin embedded or frozen sections and what conditions you would recommend for antigen retrieval and what concentration of primary antibody to use. Any advice you can give would be greatly appreciated Best Wishes

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Answer

Thank you for your enquiry regarding ab24942 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody.
I can confirm that ab24942 has been tested and characterized for ICC/IF but not for Immunohistochemistry (IHC-Fr or IHC-P). Though, it may well work in IHC but we have no data to share.
The protocol looks fine to me but I would make a few comments/suggestions:
1) Fresh samples:
It may be worth using freshly prepared and fixed samples. Try different fixatives such as ice-cold methanol or ethanol (5 mins).
2) Detection system:
Does the detection system work fine? Have you used it successfully with another primary antibody?
3) Positive control:
Brain, kidney, placenta, tonsil cells can be used as positive control. You may wish to consult this site to get some further information about the expression profile of VCAM1 in different cells/tissues:
http://www.proteinatlas.org/ENSG00000162692
I hope this will be useful for you. Should you still have any problem with this antibody after following these suggestions, then please do not hesitate to contact our Technical Department again.

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

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