- MA5-15022 - Invitrogen Antibodies GYS2 antibody

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of C2C12 (Lane 1), L6 (Lane 2), HeLa (Lane 3), A-431 (Lane 4), HEK-293 (Lane 5), Hep G2 (Lane 6), tissue extracts (30 µg lysate) of Rat Heart (Lane 7) and Mouse Skeletal Muscle (Lane 8). The blot was probed with Anti-GYS1 Monoclonal Antibody (Product # MA5-15022, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 84 kDa band corresponding to GYS1 was observed across the cell lines and tissue extracts tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of GYS1 was achieved by transfecting HeLa cells with GYS1 specific validated siRNAs (Silencer® select Product # s6372). Western blot analysis (Fig. a) was performed using whole cell extracts from the GYS1 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with GYS1 Monoclonal Antibody (Product # MA5-15022, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 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 GYS1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of C2C12 (Lane 1), L6 (Lane 2), HeLa (Lane 3), A-431 (Lane 4), HEK-293 (Lane 5), Hep G2 (Lane 6), tissue extracts (30 µg lysate) of Rat Heart (Lane 7) and Mouse Skeletal Muscle (Lane 8). The blot was probed with Anti-GYS1 Monoclonal Antibody (Product # MA5-15022, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 84 kDa band corresponding to GYS1 was observed across the cell lines and tissue extracts tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of GYS1 was performed using 70% confluent log phase HSkMs cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with GYS1 Monoclonal Antibody (Product # MA5-15022) at 1:100 dilution in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical analysis of Glycogen Synthase in paraffin-embedded human lung carcinoma using a Glycogen Synthase monoclonal antibody (Product # MA5-15022).

MA5-15022