|H M R||Endogenous||32 , 35, 36, 41||Rabbit IgG|
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.
Tropomyosin-1/3 (D17B8) Rabbit mAb detects endogenous levels of total tropomyosin-1 and/or tropomyosin-3 protein.
Human, Mouse, Rat
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu274 of human tropomyosin-1 protein.
Tropomyosin-1 (TPM1) belongs to the high molecular weight members of tropomyosin family (1,2). The protein exists in an alpha-helical coiled-coil conformation and binds multiple acting monomers in a tight manner to stabilize and regulate the actin filament (3). Tropomyosins fullfill functions in muscle and non-muscle cells. In muscle cells, tropomyosins associate with the troponin complex and play a central role in the calcium-dependent regulation of striated muscle contraction in vertebrates. In non-muscle cells, tropomyosins are implicated in the formation and stabilization of cytoskeletal actin filaments to ensure normal cellular processes (1,2). Mutations of tropomysin-1 have been reported as a cause of dilated cardiac myopathies (4). Tropomyosin-1 also functions as a tumor suppressor, and many malignant tumors demonstrate downregulation of tropomyosin-1 expression (5-8). Tropomyosin-1 is phosphorylated at Ser283 through the Erk/DAPK pathway, which promotes stress fiber formation in response to oxidative stress (9-10).
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