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Anti-GSK3B Antibody Y174
Also for GSK3B (NM_002093)
|A synthetic peptide corresponding to residues in the C-term of human GSK-3ß was used as immunogen. This antibody may detect splice isoform 2 based on sequence homology.|
||Lot dependent; please refer to CoA along with shipment
|WB, IHC, IF, FC
||WB: 1:5,000-10,000; IHC: 1:100-250; ICC: 1:100-250
|50 mM Tris-Glycine (pH 7.4), 0.15 M NaCl, 40% Glycerol, 0.01% sodium azide and 0.05% BSA.|
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Anti-GSK3 beta antibody [Y174] (TA303564) at 1/10000 dilution + A431 cell lysate.
All lanes : Anti-GSK3 beta antibody [Y174] (TA303564) at 1/2500 dilution. Lane 1 : Cell lysate prepared from MCF-7 cells; Lane 2 : Cell lysate prepared from SW480 cells. 20ug lysate/protein per lane. Donkey polyclonal IRDye 800CW at 1/15000 dilution.
IHC image of GSK3 staining in human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with TA303846, 1/200 diution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
TA303564 at a 1:100 dilution staining GSK3 beta in human colon carcinoma using Immunohistochemistry, Paraffin Embedded Tissue.TA303564 at a 1:100 dilution staining GSK3 beta in human colon carcinoma using Immunohistochemistry, Paraffin Embedded Tissue.
ICC/IF image of TA303564 stained DU145 cells. The cells were 4% formaldehyde fixed (10 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 TA303564 at 1/200 dilution overnight at +4°C. The secondary antibody (green) was DyLight488 goat anti- rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor594 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 1.43uM.
Overlay histogram showing HeLa cells stained with TA303564 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (TA303564, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1ug/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.