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Western blot
ImmunoprecipitationAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [3]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
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- Product number
- MA5-15022 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- GYS1 Monoclonal Antibody (J.18.5)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- GYS1 Monoclonal Antibody (J.18.5) detects endogenous levels of total muscle and liver glycogen synthase protein. It is not recommended to aliquot this antibody.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- J.18.5
- Vial size
- 100 µL
- Concentration
- 24 µg/mL
- Storage
- -20°C
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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.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- 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.
- 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)
- Main image
- 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)
- Main image
- Experimental details
- Immunohistochemical analysis of Glycogen Synthase in paraffin-embedded human lung carcinoma using a Glycogen Synthase monoclonal antibody (Product # MA5-15022).
