This antibody gave a positive signal in the following lysates: Human brain tissue; Human skeletal muscle tissue; Mouse brain tissue; HeLa Whole Cell; NIH 3T3 Whole Cell; K562 Whole Cell. ICC/IF: SKNSH cells.
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The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/200. Fix cells using paraformaldehyde and permeabilize with 0.5% Triton X-100/PBS before staining.
Use a concentration of 1 µg/ml. Detects a band of approximately 28 kDa (predicted molecular weight: 28 kDa).
Adapter protein implicated in the regulation of a large spectrum of both general and specialized signaling pathways. Binds to a large number of partners, usually by recognition of a phosphoserine or phosphothreonine motif. Binding generally results in the modulation of the activity of the binding partner.
Highly expressed in brain, skeletal muscle, and heart.
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Anti-14-3-3 gamma antibody 画像
Western blot - Anti-14-3-3 gamma antibody (ab102673)
All lanes : Anti-14-3-3 gamma antibody (ab102673) at 1 µg/ml
Lane 1 : Human brain tissue lysate - total protein (ab29466) Lane 2 : Human skeletal muscle tissue lysate - total protein (ab29330) Lane 3 : Brain (Mouse) Tissue Lysate Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 5 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate Lane 6 : K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution Developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 28 kDa Observed band size : 28 kDa Additional bands at : 150 kDa,40 kDa,60 kDa,90 kDa,>250 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 8 minutes
Immunocytochemistry/ Immunofluorescence - Anti-14-3-3 gamma antibody (ab102673)Image courtesy of an Abreview submitted by Dr. Kirk McManus, Univ. of Manitoba/Cancer Care MICB, Canada
ab102673 (1/200) staining 14-3-3 gamma in assynchronous HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilized with 0.5% Triton X-100/PBS and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please refer to Abreview.
ICC/IF image of ab102673 stained SKNSH cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab102673, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.