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Western blot performed from 100ng of synthetic Humanin peptide and from 50 ug of total lysates of 293 T cells transfected with plasmids encoding GFPc1 or GFP-Humanin Conditions for the Western Blot: Lysis buffer: (50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 20 mM EDTA, 50 mM NaF, 0.5% NP-40, 0.1 mM Na3VO4, 20 µg ml-1 leupeptin, 20 µg ml-1 aprotinin, 1 mM dithiothreitol and 1 mM phenylmethyl sulphonyl fluoride (PMSF)). Blocking buffer: (TRiS/Sodium chloride/buffer pH7.6) with 0.1% Tween-20, 5% skim Milk and 2% BSA). Humanin primary antibody(dilution 1/1000) and GFP antibody (dilution 1/2000) were incubated ON at 4 degrees C. Secondary rabbit HRP antibody was incubated during 1 hour at RT following by ECL detection.
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Residues 9-24 of the human Humanin protein.|
|Purification||Antigen affinity chromatography|
|Contains||0.02% sodium azide|
|Storage Conditions||Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:500|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
A gene, Humanin, encodes a short polypeptide and abolishes death of neuronal cells caused by multiple different types of familial Alzheimer's disease genes and by Abeta amyloid, without effect on death by Q79 or superoxide dismutase-1 mutants. This polype
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