|M R||Endogenous||35-45, 85-100||Rabbit|
Western blot analysis of extracts from 3T3, C2C12, and YB2/0 cells using VCAM-1 Antibody (Rodent Specific).Learn more about how we get our images.
Western blot analysis of extracts from MS-1 cells, untreated (-) or treated with Mouse Tumor Necrosis Factor-α (mTNF-α) #5178 (10 ng/ml, 16 hr; +), using VCAM-1 Antibody (Rodent Specific) (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).Learn more about how we get our images.
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length mouse VCAM-1 protein (mVCAM-1-Myc/DDK; +), using VCAM-1 Antibody (Rodent Specific).Learn more about how we get our images.
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody datasheet or product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised November 2013
Reprobing of an existing membrane is a convenient means to immunoblot for multiple proteins independently when only a limited amount of sample is available. It should be noted that for the best possible results a fresh blot is always recommended. Reprobing can be a valuable method but with each reprobing of a blot there is potential for increased background signal. Additionally, it is recommended that you verify the removal of the first antibody complex prior to reprobing so that signal attributed to binding of the new antibody is not leftover signal from the first immunoblotting experiment. This can be done by re-exposing the blot to ECL reagents and making sure there is no signal prior to adding the next primary antibody.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalently purified water.
posted June 2005
revised October 2016
Protocol Id: 10
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
VCAM-1 Antibody (Rodent Specific) recognizes endogenous levels of total VCAM-1 protein. This antibody recognizes both long and short isoforms of VCAM-1 protein.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gln203 of mouse VCAM-1 protein. Antibodies are purified by protein A and peptide affinity chromatography.
VCAM-1 (vascular cell adhesion molecule-1) is a transmembrane glycoprotein containing multiple amino-terminal extracellular Ig-like domains, a transmembrane domain, and a short carboxy-terminal cytoplasmic domain (1). Alternative splicing generates two isoforms of VCAM-1 (2). The role of VCAM-1 during infection and inflammatory diseases is well characterized. Expression of VCAM-1 is induced in endothelial cells by inflammatory cytokines including TNF-α and IL-1β (1). VCAM-1 on endothelial cells interacts with the integrin VLA-4 (α4β1) on leukocytes to mediate migration of circulating leukocytes from the blood across the endothelium and into tissues (3).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. Tween is a registered trademark of ICI Americas, Inc.
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|14694S||100 µl||$ 255.0|