Western blot analysis of extracts from various cell lines using Lu/BCAM Antibody (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
|MW (kDa)||85, 88|
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.
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
Lu/BCAM Antigody recognizes endogenous levels of total Lu/BCAM protein.Species Reactivity:
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala587 of human Lu/BCAM protein. Antibodies are purified by protein A and peptide affinity chromatography.
Lu/BCAM (CD239) is a transmembrane cell adhesion molecule belonging to the immunoglobulin superfamily (1). The protein mediates cell adhesion by binding to basal membrane laminin α5, and cell surface integrin α4β1 (2,3). The interaction between Lu/BCAM and its ligands can be activated via phosphorylation in its cytoplasmic tail by either PKA or Akt signaling (4,5). In sickle cell disease, Lu/BCAM was reported to promote abnormal red blood cell adhesion to endothelium, and thus may contribute to vaso-occlusive crisis (6). Overexpression of Lu/BCAM has also been reported in breast cancer, skin cancer, colon cancer, and ovarian cancer (7, 8). Antibody-drug conjugates targeting Lu/BCAM on tumor cells surface has been proposed as a potential approach for cancer therapy (8).
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