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, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
LCN2 (D4M8L) Rabbit mAb recognizes endogenous levels of total LCN2 protein.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ala73 of human LCN2 protein.
Lipocalin 2 (LCN2), also known as neutrophil gelatinase-associated lipocalin (NGAL), belongs to the lipocalin family. Proteins in this family exhibit considerable sequence diversity, but share a highly conserved structure composed of an eight-stranded antiparallel beta-barrel that encloses an internal ligand-binding site (1,2). LCN2 is considered as an iron transporter. LCN2 can bind bacterial siderophores and inhibits bacterial growth by iron depletion (3). It can also bind endogenous compounds like catechol in complex with iron (4). LCN2 was originally identified in neutrophils, and its expression was induced in many other tissues like kidney and liver in response to various pathologic states, such as infection, ischemia, and acute kidney injury. LCN2 is considered a biomarker for conditions like ischemic stroke, acute kidney injury, inflammatory and metabolic diseases (5-7).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. Tween is a registered trademark of ICI Americas, Inc.
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|44058S||100 µl (10 western blots)||$255.00.0|