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.
NGF Antibody detects recombinant NGF protein. It does not cross-react with other proteins.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Thr146 of human NGF.
Nerve growth factor (NGF) is a small, secreted protein and member of the neurotrophin family of growth factors that promote neuronal cell survival and differentiation (1). Producing cells release NGF that bind and activate TrkA high affinity receptors to mediate NGF-driven signaling. NGF also binds to a low affinity p75 (NTR) receptors, which belong to the death receptor family (2). Although NGF has been classically described as favoring neuron survival and differentiation, nerve growth factor can promote apoptosis in cells that contain p75 (NTR) and lack TrkA. NGF can induce neuron death in a variety of neurodegenerative conditions, including Alzheimer disease (3). Besides its neurotrophic actions, NGF has an effect on non-neuronal cells and may help mediate inflammation, angiogenesis, and stimulate breast cancer cell growth (4-6). NGF signaling is looking increasingly promising as potential drug targets for diseases.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
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|2046S||100 µl (10 western blots)||$255.00.0|