Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [1]
- Immunocytochemistry [1]
- Other assay [1]
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Validation data
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- Product number
- PA1-18285 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- ATG5 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- Reconstitute with 500 µL of distilled water. After reconstitution, store aliquots at -20ºC for a higher stability, and at 4ºC with an appropriate antibacterial agent.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 500 µg
- Concentration
- 1 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles. Glycerol (1:1) may be added for added stability.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of APG5 was performed by loading 30 µg of HEK293T whole cell lysate per well onto an SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat dry milk in TBST for 1 hour at room temperature. The membrane was probed with an APG5 polyclonal antibody (Product # PA1-18285) at a dilution of 1:200 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated goat anti-rabbit IgG secondary antibody at a dilution of 1:40,000 for 1 hour at room temperature. Detection was performed using ECL substrate. Data courtesy of the Innovators Program.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of APG5 (red) in HEK293T cells. Cells fixed with 4% formaldehyde were permeabilized and blocked with 1X PBS containing 5% BSA and 0.3% Triton X-100 for 1 hour at room temperature. Cells were probed with an APG5 polyclonal antibody (Product # PA1-18285) at a dilution of 1:20 overnight at 4°C in 1X PBS containing 1% BSA and 0.3% Triton X-100, washed with 1X PBS, and incubated with a fluorophore-conjugated goat anti-rabbit IgG secondary antibody at a dilution of 1:200 for 1 hour at room temperature. Nuclei (blue) were stained with DAPI. Images were taken at 20X magnification. Data courtesy of the Innovators Program.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunoprecipitation of APG5 was performed on HEK293T cells. Antigen-antibody complexes were formed by incubating 500 µg of HEK293T whole cell lysate (in 500 µL volume) with 5 µL of an APG5 Antibody (Product # PA1-18285) overnight at 4°C. The immune complexes were captured on 30 µL of protein G agarose, washed extensively, and eluted with 6X non-reducing Laemmli buffer. Samples were resolved on a 12% SDS-PAGE, transferred to a PVDF membrane, and blocked with 5% milk in TBST for 1 hour at room temperature. The membrane was probed with an APG5 polyclonal antibody (Product # PA1-18285) at a dilution of 1:200 overnight at 4øC, washed in TBST, and probed with an HRP-conjugated goat anti-rabbit IgG secondary antibody at a dilution of 1:40,000 for 1 hour at room temperature. Chemiluminescent detection was performed using ECL substrate. Data courtesy of the Innovators Program.