|H M R Mk||Endogenous||55||Rabbit IgG|
Western blot analysis of extracts from human prostate tissue and SK-MEL-5 and Malme-3M cells using SNX17 (E8F8Y) Rabbit mAb (upper) and β-Tubulin (D2N5G) Rabbit mAb #15115 (lower).Learn more about how we get our images.
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.
SNX17 (E8F8Y) Rabbit mAb recognizes endogenous levels of total SNX17 protein. Non-specific bands may be detected above 120 kDa in some cases.
Human, Mouse, Rat, Monkey
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Asn54 of human SNX17 protein.
Sorting nexins are a family of cytoplasmic proteins characterized by the presence of a phosphatidylinositol 3-phosphate (PI3P) binding phox (PX) domain. This binding occurs mainly in the early endosome and allows for trafficking of the bound protein to either a degradative or recycling pathway (1).
Sorting nexin-17, or SNX17, has been shown to preferentially drive trafficking of integrins, receptors, and a variety of other proteins away from degradative pathways (1). In addition to PX domain interactions, SNX17 also binds the NPxY motif on the cytoplasmic tails of lipoprotein receptors via its FERM domain (protein 4.1, ezrin, radixin and moesin). Some of these proteins include the low density lipoprotein receptor-related protein 1 (LRP1) and apolipoprotein E receptor 2 (ApoER2) (3,4). LRP1 is known to bind APP, regulating its processing and causing an increase in Aβ production, a known risk factor for AD. By binding APP in addition to LRP1, SNX17 recycles both proteins to the plasma membrane, maintaining normal cell surface levels of each (3). SNX17 acts similarly with ApoER2, facilitating trafficking and increasing recycling to the plasma membrane. This assists in regulating the binding of ApoER2 and reelin, an interaction that is known to be important for neuronal migration and the formation of brain structures in early development, as well as synaptic function, learning, and memory in the adult brain (4). Through these and other interactions, SNX17 has been shown to have a potential role in a wide variety of neuronal pathways and diseases.
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|75885S||100 µl (10 western blots)||$ 255.0|