|H M R Mk||Endogenous||95||Rabbit|
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.
Nod1 Antibody detects endogenous levels of Nod1 protein.
Human, Mouse, Rat, Monkey
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of Nod1 protein. Antibodies were purified by protein A and peptide affinity chromatography.
Nod1/CARD4 is a cytosolic protein structually related to Apaf-1 and plant drug-resistance proteins that has been implicated in apoptosis and inflammatory responses to certain pathogenic bacteria (1-3). It contains an amino-terminal caspase recruitment domain (CARD) that is linked to a central nucleotide-binding domain (NBD; also known as a NOD domain) and is followed by carboxy-terminal leucine-rich repeats (LRR) (1). Like Apaf-1, Nod1 induces apoptosis by a CARD/NBD-dependent activation of caspase-9 (1). The primary function of Nod1 is thought to be as a sensor for certain pathogenic microbes and triggering inflammatory responses including the activation of NF-κB and JNK pathways (4-6). The LRR of Nod1 appears to be involved in recognition of microbial components and the CARD domain induces NF-κB activation in cooperation with the CARD containing kinase, RICK/RIP2/CARDIAK (1,5,6). Mutations in Nod1 have been linked increased susceptibility to asthma (7) and inflammatory bowel disease (8).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
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