Anti-VCAM1 antibody (ab115135)
Key features and details
- Rabbit polyclonal to VCAM1
- Suitable for: WB, ICC/IF
- Reacts with: Mouse, Rat
- Isotype: IgG
Overview
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Product name
Anti-VCAM1 antibody
See all VCAM1 primary antibodies -
Description
Rabbit polyclonal to VCAM1 -
Host species
Rabbit -
Tested applications
Suitable for: WB, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat
Predicted to work with: Horse, Macaque monkey
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Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Mouse Spleen, Rat Spleen, Mouse Thymus, Rat Thymus, and Mouse Lung tissue lysates; LADMAC and LPS treated bEnd.3 cell lysates; ICC/IF: NIH-3T3 cells.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
Our Abpromise guarantee covers the use of ab115135 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 a concentration of 1 µg/ml. Detects a band of approximately 100 kDa (predicted molecular weight: 81 kDa). Stimulation may be required to allow detection of the target protein due to low levels of endogenous expression in some samples. Please see images below for recommended treatment conditions and positive controls.
ICC/IF Use a concentration of 5 µg/ml. Target
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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
modificationsSialoglycoprotein. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 22329 Mouse
- Entrez Gene: 25361 Rat
- SwissProt: P29533 Mouse
- SwissProt: P29534 Rat
- Unigene: 440909 Mouse
- Unigene: 76649 Mouse
- Unigene: 11267 Rat
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Alternative names
- CD106 antibody
- CD106 Antigen antibody
- INCAM 100 antibody
see all
Images
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Western blot - Anti-VCAM1 antibody (ab115135)All lanes : Anti-VCAM1 antibody (ab115135) at 1/1000 dilution
Lane 1 : LADMAC (Mouse bone marrow monocyte macrophage) whole cell lysate
Lane 2 : bEnd.3 (Mouse brain endothelioma) whole cell lysate
Lane 3 : bEnd.3 (Mouse brain endothelioma) 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?
Exposure time: 3 minutesRabbit monoclonal [EPR16891] to GAPDH (ab181602) used as loading control.
Blocking/Diluting buffer: 5% NFDM/TBST.
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Immunocytochemistry/ Immunofluorescence - Anti-VCAM1 antibody (ab115135)ICC/IF image of ab115135 stained NIH-3T3 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab115135 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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Western blot - Anti-VCAM1 antibody (ab115135)All lanes : Anti-VCAM1 antibody (ab115135) at 1 µg/ml
Lane 1 : Spleen (Mouse) Tissue Lysate
Lane 2 : Spleen (Rat) Tissue Lysate
Lane 3 : Thymus (Mouse) Tissue Lysate
Lane 4 : Thymus (Rat) Tissue Lysate
Lane 5 :Mouse lung normal tissue lysate - total protein (ab29297)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 81 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?
Additional bands at: 62 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 4 minutes
VCAM1 contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.
Protocols
Datasheets and documents
References (0)
ab115135 has not yet been referenced specifically in any publications.
Images
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Western blot - Anti-VCAM1 antibody (ab115135)All lanes : Anti-VCAM1 antibody (ab115135) at 1/1000 dilution
Lane 1 : LADMAC (Mouse bone marrow monocyte macrophage) whole cell lysate
Lane 2 : bEnd.3 (Mouse brain endothelioma) whole cell lysate
Lane 3 : bEnd.3 (Mouse brain endothelioma) 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?
Exposure time: 3 minutesRabbit monoclonal [EPR16891] to GAPDH (ab181602) used as loading control.
Blocking/Diluting buffer: 5% NFDM/TBST.
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Immunocytochemistry/ Immunofluorescence - Anti-VCAM1 antibody (ab115135)
ICC/IF image of ab115135 stained NIH-3T3 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab115135 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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Western blot - Anti-VCAM1 antibody (ab115135)All lanes : Anti-VCAM1 antibody (ab115135) at 1 µg/ml
Lane 1 : Spleen (Mouse) Tissue Lysate
Lane 2 : Spleen (Rat) Tissue Lysate
Lane 3 : Thymus (Mouse) Tissue Lysate
Lane 4 : Thymus (Rat) Tissue Lysate
Lane 5 :Mouse lung normal tissue lysate - total protein (ab29297)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 81 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?
Additional bands at: 62 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 4 minutes
VCAM1 contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.
