Western blot analysis of extracts from SU-DHL-4 and SH-SY5Y cells using Drebrin-like protein/DBNL Antibody (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from human brain cortex using Drebrin-like protein/DBNL Antibody (upper) and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower).
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
Drebrin-like protein/DBNL Antibody recognizes endogenous levels of total Drebrin-like protein/DBNL.Species Reactivity:
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly147 of human Drebrin-like protein/DBNL. Antibodies are purified by peptide affinity chromatography.
Drebrin-like protein/DBNL is an actin-binding protein that expresses in the developing cerebral cortex and contributes to the formation and morphology of processes in the multipolar neurons. The expression of Drebrin-like protein increases the levels of N-cadherin, promoting intracellular trafficking and cell migration. High expression of N-cadherin can rescue deficits in cell migration induced by a defective Drebrin-like protein in neurons (1,2). Drebrin-like protein directly interacts with Cobl, forming a complex found in Purkinje cell bodies and dendritic arbors in the cerebellum (3). Severe impairments in the recycling of synaptic vesicles in hippocampal neurons and behavioral anomalies occur in mice deficient of Drebrin-like protein (4). The phosphorylation of Drebrin-like protein at Ser269/Thr291 promotes the localization in the cortical plate of the Golgi apparatus, facilitating the correct intracellular protein trafficking (5). Drebrin-like protein participates in regulatory network signaling by the activation of β-adrenergic receptors during the proliferation of cardiac fibroblasts, which is associated with the extracellular activation of ERK (4). Drebrin-like protein plays a role in cell migration, the extension of neuronal processes and plasticity of dendrites, and is required for the polymerization of actin at immunological synapses (IS), for CXCR4 recruitment to IS, and activation of T-cells (5-7).
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