|H M R||Endogenous||55||Mouse IgG1|
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.
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 June 2016
Protocol Id: 262
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.
CYP1A2 (D2V7S) Mouse mAb recognizes endogenous levels of total CYP1A2 protein. This antibody does not cross-react with CYP1A1 protein.
Human, Mouse, Rat
Monoclonal antibody is produced by immunizing animals with 3-methylcholanthrene-induced rat liver microsomal fractions.
Cytochrome P450 (CYP) is a family of enzymes that contain a heme group (1). These enzymes, when reduced and bound by carbon monoxide, maximally absorb light of 450 nm (1). Type I cytochrome P450s are found in mitochondria and function in the biosynthesis of essential molecules (1). Type II cytochrome P450s are found in endoplasmic reticulum (1). Some type II cytochrome P450s play a role in the biosynthesis of essential molecules whereas others metabolize xenobiotics (1). Research studies show that cytochrome P450s form various heteromeric complexes with other members of the P450 family influencing their catalytic activities (2-4). CYP1A2 is in the endoplasmic reticulum of hepatocytes and responsible for the breakdown of a variety of xenobiotic substances and bioactivation of carcinogens (2, 5). CYP1 enzymes, including CYP1A2, have been implicated in smoking-related osteoporosis (6). A meta-analysis shows that a particular polymorphism in CYP1A2 is potentially linked to increased cancer risk (5).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. Tween is a registered trademark of ICI Americas, Inc.
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|14719S||100 µl (10 western blots)||$ 255.0|