Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-VCAM1 antibody [EPR5038(2)] - BSA and Azide free (ab249790)

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Western blot - Anti-VCAM1 antibody [EPR5038(2)] - BSA and Azide free (ab249790)
  • Western blot - Anti-VCAM1 antibody [EPR5038(2)] - BSA and Azide free (ab249790)
  • Western blot - Anti-VCAM1 antibody [EPR5038(2)] - BSA and Azide free (ab249790)
  • Western blot - Anti-VCAM1 antibody [EPR5038(2)] - BSA and Azide free (ab249790)
  • Immunoprecipitation - Anti-VCAM1 antibody [EPR5038(2)] - BSA and Azide free (ab249790)
  • Anti-VCAM1 antibody [EPR5038(2)] - BSA and Azide free (ab249790)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR5038(2)] to VCAM1 - BSA and Azide free
  • Suitable for: WB, IP
  • Knockout validated
  • Reacts with: Mouse, Human

Conjugates logo Related conjugates and formulations

Unconjugated

Overview

  • Product name

    Anti-VCAM1 antibody [EPR5038(2)] - BSA and Azide free
    See all VCAM1 primary antibodies

  • Description

    Rabbit monoclonal [EPR5038(2)] to VCAM1 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IPmore details
    Unsuitable for: Flow Cyt,ICC/IF or IHC-P

  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Wild-type A549 TNF-a treated (10 ng/mL, 16h) cell lysate

  • General notes

    ab249790 is the carrier-free version of ab174279.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab249790 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).
IP
Use at an assay dependent concentration.
Notes
WB
Use at an assay dependent concentration. Detects a band of approximately 100 kDa (predicted molecular weight: 81 kDa).
IP
Use at an assay dependent concentration.
Application notes
Is unsuitable for Flow Cyt,ICC/IF or IHC-P.

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

    see all

  • 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

  • Western blot - Anti-VCAM1 antibody [EPR5038(2)] - BSA and Azide free (ab249790)
    Western blot - Anti-VCAM1 antibody [EPR5038(2)] - BSA and Azide free (ab249790)
    All lanes : Anti-VCAM1 antibody [EPR5038(2)] (ab174279) at 1/1000 dilution

    Lane 1 : Wild-type A549 cell lysate
    Lane 2 : Wild-type A549 TNF-a treated (10 ng/mL, 16h) cell lysate
    Lane 3 : VCAM1 knockout A549 cell lysate
    Lane 4 : VCAM1 knockout A549 TNF-a treated (10 ng/mL, 16h) cell lysate

    Lysates/proteins at 30 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 81 kDa
    Observed band size: 105 kDa why is the actual band size different from the predicted?


    Exposure time: 20 seconds


    ab174279 was shown to react with VCAM1 in wild-type A549 cells in western blot. Loss of signal was observed when VCAM1 knockout sample was used. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab174279 overnight at 4 °C at a 1 in 1000 dilution. Blots were incubated with HRP conjugated Goat anti-Rabbit (H+L) secondary antibody at 1 in 5000 for 1 hour at room temperature before development with Optiblot ECL reagent (ab133456) and imaging.

  • Western blot - Anti-VCAM1 antibody [EPR5038(2)] - BSA and Azide free (ab249790)
    Western blot - Anti-VCAM1 antibody [EPR5038(2)] - BSA and Azide free (ab249790)
    All lanes : Anti-VCAM1 antibody [EPR5038(2)] (ab174279) at 1/1000 dilution

    Lane 1 : HUVEC (Human umbilical vein endothelial cell) whole cell lysate
    Lane 2 : HUVEC treated with 10 ng/ml TNF-a for 16 h, whole cell lysate
    Lane 3 : bEND.3 (Mouse brain endothelioma) whole cell lysate
    Lane 4 : bEND.3 treated with 10 µg/ml LPS for 24 h, whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

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



    This data was developed using ab174279, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-VCAM1 antibody [EPR5038(2)] - BSA and Azide free (ab249790)
    Western blot - Anti-VCAM1 antibody [EPR5038(2)] - BSA and Azide free (ab249790)
    All lanes : Anti-VCAM1 antibody [EPR5038(2)] (ab174279) at 1/1000 dilution

    Lane 1 : Hut-78 (Human Sezary syndrome cutaneous T lymphocyte) whole cell lysate
    Lane 2 : SK-OV-3 (Human ovarian cancer epithelial cell) whole cell lysate
    Lane 3 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
    Lane 4 : J774A.1 (Mouse reticulum cell sarcoma monocyte macrophage) whole cell lysate
    Lane 5 : LADMAC (Mouse bone marrow monocyte macrophage) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

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


    Exposure time: 15 seconds


    This data was developed using ab174279, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-VCAM1 antibody [EPR5038(2)] - BSA and Azide free (ab249790)
    Western blot - Anti-VCAM1 antibody [EPR5038(2)] - BSA and Azide free (ab249790)
    All lanes : Anti-VCAM1 antibody [EPR5038(2)] (ab174279) at 1/1000 dilution

    Lane 1 : Human fetal kidney lysate
    Lane 2 : HuT-78 lysate
    Lane 3 : NIH3T3 lysate

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 81 kDa



    This data was developed using ab174279, the same antibody clone in a different buffer formulation.

  • Immunoprecipitation - Anti-VCAM1 antibody [EPR5038(2)] - BSA and Azide free (ab249790)
    Immunoprecipitation - Anti-VCAM1 antibody [EPR5038(2)] - BSA and Azide free (ab249790)

    This data was developed using ab174279, the same antibody clone in a different buffer formulation.

    Immunoprecipitation. ab174279 at 1/1000 labeling VCAM1 immunoprecipitated from NIH3T3 cell lysate using ab174279 at 1/10.

  • Anti-VCAM1 antibody [EPR5038(2)] - BSA and Azide free (ab249790)
    Anti-VCAM1 antibody [EPR5038(2)] - BSA and Azide free (ab249790)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (0)

Publishing research using ab249790? Please let us know so that we can cite the reference in this datasheet.

ab249790 has not yet been referenced specifically in any publications.

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