Recombinant
RabMAb

Anti-VCAM1 antibody [EPR5047] (ab134047)

Overview

  • Product name
    Anti-VCAM1 antibody [EPR5047]
    See all VCAM1 primary antibodies
  • Description
    Rabbit monoclonal [EPR5047] to VCAM1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IF, IHC-Frmore 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. FC: K562 cells
  • 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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab134047 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 1/2000 - 1/10000. Detects a band of approximately 100 kDa (predicted molecular weight: 81 kDa).Can be blocked with VCAM1 peptide (ab156177).
IP 1/40.
IHC-P 1/500 - 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocol.

Flow Cyt 1/40.
ICC/IF 1/250.
IHC-Fr 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

  • 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.

  • 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.

  • All lanes : Anti-VCAM1 antibody [EPR5047] (ab134047) at 1/10000 dilution (purified)

    Lane 1 : Mouse kidney
    Lane 2 : Rat spleen

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 81 kDa
    Observed band size: 100 kDa (why is the actual band size different from the predicted?)



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • 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.

  • 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.

  • Anti-VCAM1 antibody [EPR5047] (ab134047) at 1/2000 dilution (purified) + Rat kidney at 20 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 81 kDa
    Observed band size: 100 kDa (why is the actual band size different from the predicted?)



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Anti-VCAM1 antibody [EPR5047] (ab134047) at 1/10000 dilution (purified) + NIH/3T3 at 10 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 81 kDa
    Observed band size: 100 kDa (why is the actual band size different from the predicted?)



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • 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

  • Anti-VCAM1 antibody [EPR5047] (ab134047) at 1/10000 dilution (purified) + Human fetal liver at 20 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 81 kDa
    Observed band size: 100 kDa (why is the actual band size different from the predicted?)



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Anti-VCAM1 antibody [EPR5047] (ab134047) at 1/10000 dilution (purified) + HuT-78 cell lysate at 10 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 81 kDa
    Observed band size: 100 kDa (why is the actual band size different from the predicted?)



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • IHC image of VCAM1 staining in Mouse spleen formalin fixed paraffin embedded tissue section, performed on a Leica Bond

    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.

  • All lanes : Anti-VCAM1 antibody [EPR5047] (ab134047) at 1/1000 dilution (unpurified)

    Lane 1 : Human fetal liver lysate
    Lane 2 : HuT 78 lysate
    Lane 3 : NIH 3T3 lysate
    Lane 4 : Mouse brain lysate
    Lane 5 : Mouse kidney lysate
    Lane 6 : Mouse spleen lysate
    Lane 7 : Rat brain lysate
    Lane 8 : Rat kidney lysate
    Lane 9 : Rat spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 81 kDa



    Secondary antibody - goat anti-rabbit HRP (ab6721)

References

This product has been referenced in:
  • Lv P  et al. Apolipoprotein C-III in the high-density lipoprotein proteome of cerebral lacunar infarction patients impairs its anti-inflammatory function. Int J Mol Med 41:61-68 (2018). Read more (PubMed: 29115584) »
  • You S  et al. An Aza resveratrol-chalcone derivative 6b protects mice against diabetic cardiomyopathy by alleviating inflammation and oxidative stress. J Cell Mol Med 22:1931-1943 (2018). Read more (PubMed: 29327811) »

See all 32 Publications for this product

Customer reviews and Q&As

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - other (COS cells)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
20 µg
Specification
COS cells
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Oct 27 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Human pluripotent SC derived epicardial like cells)
Permeabilization
Yes - Saponin
Specification
Human pluripotent SC derived epicardial like cells
Blocking step
Serum as blocking agent for 15 minute(s) · Concentration: 4% · Temperature: 4°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Oct 24 2016

Anti-VCAM1 antibody [EPR5047] (ab134047) is a rabbit IgG. Unlike mice, rabbits have only one IgG subclass.

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (vessels)
Antigen retrieval step
Other
Permeabilization
No
Specification
vessels
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Fixative
zinc buffered formalin
Username

Abcam user community

Verified customer

Submitted Feb 23 2016

Application
Immunocytochemistry
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Sample
Mouse Cultured Cells (cardiomyocyte differentiation)
Specification
cardiomyocyte differentiation
Permeabilization
Yes - 0,02% TritonX-100 (in PBS)
Fixative
Paraformaldehyde
Username

Mr. Tamas Bellak

Verified customer

Submitted Jan 30 2015

Application
Immunohistochemistry (Frozen sections)
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Sample
Mouse Tissue sections (Aorta)
Specification
Aorta
Permeabilization
No
Fixative
Methanol
Username

Mrs. Anna Maksin

Verified customer

Submitted Apr 01 2014

I agree that the reason for the ˜100 kDa band is that the VCAM1 is a glycoprotein, so the actual band size detected by the antibody is higher than the theoretical size.
This reference also suggests the same:
http://www.ncbi.nlm.nih.gov/pubmed...

Read More

Merci de nous avoir contactés.

La séquence immunogène d’ab134047 est xxxxxxx qui correspond aux acides aminés xxxxxxxxx de la région C-terminale de la protéine chez la souris. Cependant, cette s...

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