MA5-15597

antibody from Invitrogen Antibodies

Targeting: GSK3B

Western blot (Supportive data in Antibodypedia)

ELISA (Recommended by provider)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-15597

Provider

Invitrogen Antibodies

Product name

GSK3B Monoclonal Antibody (3D10)

Antibody type

Monoclonal

Antigen

Purifed from natural sources

Reactivity

Human, Mouse, Rat

Host

Mouse

Isotype

IgG

Antibody clone number

3D10

Vial size

100 µL

Concentration

Conc. Not Determined

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

References

Submitted references

Proteomic characterization of post-mortem human brain tissue following ultracentrifugation-based subcellular fractionation.

Kandigian SE, Ethier EC, Kitchen RR, Lam TT, Arnold SE, Carlyle BC
Brain communications 2022;4(3):fcac103

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of GSK3B was achieved by transfecting MCF7 cells with GSK3B specific validated siRNAs (Silencer® select Product # S6240). Western blot analysis (Fig. a) was performed using whole cell extracts from the GSK3B knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with GSK3B Monoclonal Antibody (Product # MA5-15597, 1:500 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to GSK3B.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on whole cell extracts of MCF7 (Lane 1), MDA-MB-231 (Lane 2), HeLa (Lane 3), Neuro-2a (Lane 4), U-87 MG (Lane 5), A-431 ( Lane 6) and NIH/3T3 (Lane 7). The blot was probed with Anti-GSK3B antibody (Product # MA5-15597, 1:500 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). A 47 kDa band corresponding to 4EBP1 was observed across the cell lines tested.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of NIH/3T3 (left) and U251 (right) cells using GSK3B monoclonal antibody (Product # MA5-15597) (Green). Blue: DRAQ5 fluorescent DNA dye. Red: actin filaments have been labeled with phalloidin.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of paraffin-embedded human lung cancer (left) and breast cancer tissues (right) using GSK3B monoclonal antibody (Product # MA5-15597) followed with DAB staining.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometric analysis of HeLa cells using GSK3B monoclonal antibody (Product # MA5-15597) (green) and negative control (purple).

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry of angular gyrus sections with anti-GSK3beta shows a trend towards increased nuclear GSK3beta in controls . ( A ) Representative images show increased colocalization of GSK3beta signal with nuclear DAPI staining. Subjective visual analysis suggests increased presence of nuclear speckles in Control samples compared with AD. ( B ) Plot showing Mander's M2 overlap coefficient for each individual image shows an enrichment for increased nuclear overlap of GSK3beta staining with DAPI. ( C ) The mean Mander's M2 coefficient for each subject shows a trend (Student's t -test, T = -2.18, P = 0.07) towards increased nuclear overlap of GSK3beta in controls compared with AD.