MA5-15109

antibody from Invitrogen Antibodies

Targeting: GSK3B

Immunoprecipitation

Immunohistochemistry

Antibody data

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Validation data

GSK3B protein structure - MA5-15109 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of GSK3B was performed with 10 µg of A549 cells transfected with Transfection Reagent alone (Lane 1), 100nM Non-Targeting control siRNA (Lane 2), or 100nM siRNA against GSK3B (Lane 3). Proteins were resolved using a NuPAGE® Novex 4-12% Bis-Tris Gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002), and a protein size ladder. Proteins were wet transferred to a Pierce Nitrocellulose Membrane (Product # 88025) OR Pierce PVDF Membrane (Product # 88518) and blocked with Pierce Starting Block T20 (PBS) Blocking Buffer (Product # 37539) for 1 hour at room temperature. GSK3B was detected at ~ 46 kDa using GSK3B Rabbit monoclonal antibody (Product # MA5-15109) diluted in Pierce Starting Block T20 (PBS) Blocking Buffer 4°C overnight on a rocking platform. Pierce Goat Anti-Rabbit (Product # 31461) HRP-Conjugated Antibodies at a 1:2500 dilution were used and chemiluminescent detection was performed using Pierce Supersignal West Dura Maximum Sensitivity Substrate (Product # 37071). Relative density of the bands normalized to Cyclophilin B (21 kDa). GSK3B Antibody (Product # MA5-15109) confirms silencing of GSK3B expression.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of GSK3B was performed with 10 µg of A549 cells transfected with Transfection Reagent alone (Lane 1), 100nM Non-Targeting control siRNA (Lane 2), or 100nM siRNA against GSK3B (Lane 3). Proteins were resolved using a NuPAGE® Novex 4-12% Bis-Tris Gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002), and a protein size ladder. Proteins were wet transferred to a Pierce Nitrocellulose Membrane (Product # 88025) OR Pierce PVDF Membrane (Product # 88518) and blocked with Pierce Starting Block T20 (PBS) Blocking Buffer (Product # 37539) for 1 hour at room temperature. GSK3B was detected at ~ 46 kDa using GSK3B Rabbit monoclonal antibody (Product # MA5-15109) diluted in Pierce Starting Block T20 (PBS) Blocking Buffer 4°C overnight on a rocking platform. Pierce Goat Anti-Rabbit (Product # 31461) HRP-Conjugated Antibodies at a 1:2500 dilution were used and chemiluminescent detection was performed using Pierce Supersignal West Dura Maximum Sensitivity Substrate (Product # 37071). Relative density of the bands normalized to Cyclophilin B (21 kDa). GSK3B Antibody (Product # MA5-15109) confirms silencing of GSK3B expression.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 1 Downregulation of GSK-3beta activity by HDAC6. a Neuro2a mapped for total GSK-3beta shows their unaltered levels upon HDAC6 ZnF UBP. b Inhibitory phosphorylation of GSK-3beta at Ser9 increases upon HDAC6 ZnF UBP treatment. The enlarged image shows the elevated level of pGSK-3beta compared to neuro2a cell control. c Quantification of mean fluorescence intensity for GSK-3beta in cell control and HDAC6 ZnF UBP treated cells showed non-significant difference while the levels of pGSK-3beta were significantly increased upon HDAC6 ZnF UBP treatment in comparison to cell control

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2. Melatonin Altered GSK3beta Expression but Not Activation. A: In AD, Tau becomes hyperphosphorylated and subsequently deposits as aggregates in neurons. OA has been proven as a model to induce Tau phosphorylation via PP2A inhibition and enhancement of overall kinase activity. Melatonin has been proposed to block the GSK3beta-mRNA expression and total protein level. B: Melatonin reduced the total-GSK3beta mRNA expression level on Neuro2A cells upon OA treatment by qRT-PCR. C and D: OA has induced the Ser 9 phosphorylation of GSK3beta due to the overall enhancement of global phosphorylation, whereas the level of total GSK3beta decreased slightly. E: Western blot densitometric quantification showed a decreased level of total GSK3beta in Melatonin-treated Neuro2A cells. F: Immunofluorescence study depicted the altered level of only total GSK3beta protein, but GSK3beta (phopsho Ser9) remained invariant. G and H: The relative fluorescence level was plotted for Melatonin and OA treated group alone and together. * corresponds to p-values of test groups compared with untreated control (*p < 0.05; **p